Compositions comprising Paulownia tomentosa wood extracts and uses thereof

ABSTRACT

Provided are compositions comprising an extract of  Paulownia tomentosa  wood and methods of use thereof.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a continuation-in-part of U.S. application Ser. No.12/859,323, filed Aug. 19, 2010 now abandoned.

FIELD OF INVENTION

The present invention relates to compositions comprising plant extractsfor use on skin. More specifically, it relates to compositionscomprising extracts of Paulownia tomentosa wood for a variety of uses onthe skin.

DESCRIPTION OF RELATED ART

Paulownia is a genus of plants native to Asia which has spread graduallyto Europe and the USA. Species from the Paulownia genus are generallyconsidered ornamental trees with wide craft and ecological applicationsfor their wood. For example, the wood of such trees is popular formaking soundboards of stringed instruments in Japan, China, and Korea.It is also popular with timber merchants for use in making furniture.Paulownia trees have ecological uses and are consideredphyto-remidiators, that is, they can process industrial contaminantsthrough their vascular system to help clean and reclaim land.

In Japan, Paulownia is called kiri which refers specifically to onespecies, Paulownia tomentosa, also called “Princess Tree.” Other nameswhich are commonly used are “empress tree”, “Foxglove Tree”, “RoyalPaulownia”, “Pao tong” (in China) and “Odong-Namoo” (in Korea). Thescientific name is “Paulownia tomentosa” with a number of synonymsreported in various literature, i.e. “Paulownia imperialis”, “Paulowniarecurva”, and “Bignonia tomentosa”. Paulownia tomentosa belongs to thefamily “Paulowniaceae” or sometimes refer to “Scrophulariaceae”. TheUnited States Department of Agriculture (plants.USDA.gov) Plant databaseidentifies Princess tree by a unique symbol “PATO2”, with Paulowniatomentosa and Paulownia imperialis as synonym names.

The flower oil of Paulownia tomentosa is well studied and found to bericher in aroma as compared to other species. A number of bioactivitiesare associated with extracts of various parts of Paulownia, e.g.anti-cancer components from flower extract, anthelminthic activity fromnon-specified extract, antibacterial activities from fruit and flowerextracts, antioxidant activity from flower extract, and anti-viralproperties from stem bark. Leaf extracts of Paulownia are described forhair growth and hair promoting properties.

The present invention relates to applicant's discovery that extracts ofPaulownia tomentosa wood are beneficial for use in compositions on skin.For example, applicants have discovered that such compositions tend toexhibit significant and unexpected properties for skin including skinlightening, improving signs of aging, and reducing inflammation.

SUMMARY OF THE INVENTION

In one aspect, the present invention is directed to compositionscomprising an extract of Paulownia tomentosa wood and a carrier.

In another aspect, the present invention is directed to methods oflightening the skin comprising the step of applying to skin in need ofskin lightening treatment an extract of Paulownia tomentosa wood.

In another aspect, the present invention is directed to methods ofimproving a sign of aging in skin comprising the step of applying toskin in need of improving the signs of aging an extract of Paulowniatomentosa wood.

In yet another aspect, the present invention is directed to methods ofreducing skin inflammation comprising the step of applying to a skin inneed of reducing skin inflammation an extract of Paulownia tomentosawood.

DESCRIPTION OF THE INVENTION

As noted above, applicants have discovered unexpectedly that extracts ofthe wood of Paulownia tomentosa may be used in compositions, preferablyskin care compositions, and for methods of use including, but notlimited to, skin lightening, improving signs of aging, and reducinginflammation.

As used herein, the term “lightening the skin” refers generally tolightening, brightening, whitening, and/or evening of the skin tone,skin color, and/or shade of skin, and/or to the reduction in sallowness,and/or to the lightening and/or fading of hyperpigmented marks and/orlesions including, but not limited to, pigmented spots, melanin spots,age spots, sun spots, senile lentigos, freckles, lentigos simplex,pigmented solar keratosis, seborrhoeic keratosis, melasma, acne marks,post-inflammatory hyperpigmentation, lentigines, ephelides, combinationsof two or more thereof and the like. In certain embodiments, “lighteningthe skin” also refers to increased skin radiance, glow, translucencyand/or luminescence and/or obtaining a more radiant, glowing,translucent or luminous skin tone appearance or a less yellow or sallowskin tone. In certain preferred embodiments, “lightening the skin”refers to lightening and evening the skin tone, increasing skin radianceand/or lightening age spots.

As used herein, the term “skin in need of skin lightening treatment”refers generally to skin that exhibits one or more property selectedfrom the group consisting of: skin having a measured Individual TypologyAngle (ITA) value below 41 as determined per the COLIPA GUIDELINE:GUIDELINE FOR THE COLORIMETRIC DETERMINATION OF SKIN COLOUR TYPING ANDPREDICTION OF THE MINIMAL ERYTHEMAL DOSE (MED) WITHOUT UV EXPOSUREpublished in 2007, which is incorporated herein by reference and furtherdescribed below, darkened and/or sallow skin, including skin darkened byUV, skin with uneven skin tone, or skin with one or more hyperpigmentedmarks and/or lesions including, but not limited to, pigmented spots,melanin spots, age spots, sun spots, senile lentigos, freckles, lentigossimplex, pigmented solar keratosis, seborrhoeic keratosis, melasma, acnemarks, post-inflammatory hyperpigmentation, lentigines, ephelides,combinations of two or more thereof and the like. In the COLIPAguidelines, skin color is defined function of the ITA value as: verylight skin >55; Light skin 41-55, Intermediate 28-41, and Tan skin <28.In certain preferred embodiments, “skin in need of skin lightening”refers to individuals with a skin having an ITA value of less than 41,such as about 40 or less, about 35 or less, about 30 or less, or morepreferably about 28 or less. In certain other preferred embodiments, thepresent invention is directed to compositions and methods for use onskin in need of skin lightening treatment selected from sallow and/ordarkened skin. In certain other preferred embodiments, the presentinvention is directed to compositions and methods for use on skin inneed of skin lightening treatment selected from the group consisting ofage spots, freckles, marks left after acne, and combinations of two ormore thereof.

As used herein, “skin in need of improving the signs of aging” means askin that is, but not limited to, sagging, loose, lax, rough, wrinkly,thinned, uneven. Improving the signs of aging means improving thefirmness of the skin, improving the texture of the skin, improving theappearance of wrinkles in skin, improving the skin tone or the treatmentof external aggressions in skin.

As used herein, “improving the firmness of skin” means the enhancing ofthe firmness or elasticity of the skin, preventing the loss of firmnessor elasticity of skin, or preventing or treating sagging, lax and looseskin. The firmness or elasticity of the skin can be measured by use of acutometer. See Handbook of Non-Invasive Methods and the Skin, eds. J.Serup, G. Jemec & G. Grove, Chapter 66.1 (2006). The loss of skinelasticity or firmness may be a result of a number of factors, includingbut not limited to aging, environmental damage, or the result of anapplication of a cosmetic to the skin.

As used herein, “improving the texture of skin” means the smoothing ofthe surface of the skin to remove either bumps or crevasses on the skinsurface.

As used herein, “improving the appearance of wrinkles in skin” meanspreventing, retarding, arresting, or reversing the process of wrinkleand fine line formation in skin.

As used herein, “treatment of external aggressions in skin” means thereduction or prevention of the damage from external aggressions in skin.Examples of external aggressions include, but are not limited to, damageto the skin from the use of cleansers (e.g., topical cleanserscontaining surfactants), make-up, shaving as well as environmentaldamage such as from UV light (e.g., sundamage from sunlight or damagefrom non-natural sources such as UV lamps and solar simulators), ozone,exhaust, pollution, chlorine and chlorine containing compounds, andcigarette smoke. Effects of external aggressions on the skin include,but are not limited to, oxidative and/or nitrosative damage to andmodifications on lipids, carbohydrates, peptides, proteins, nucleicacids, and vitamins. Effects of external aggressions on the skin alsoinclude, but are not limited to, loss of cell viability, loss oralteration of cell functions, and changes in gene and/or proteinexpression.

As used herein, “improving the skin tone” means the lightening of theappearance of the skin (e.g., lightening pigmented marks or lesions,reducing skin sallowness, and/or evening the color of the skin).

As used herein, “skin in need of reducing skin inflammation” means askin exhibiting redness or erythema, edema, or being reactive orsensitive to external elements. External elements include, but are notlimited to, sun rays (UV, visible, IR), microorganisms, atmosphericpollutants such as ozone, exhaust pollutants, chlorine and chlorinegenerating compounds, cigarette smoke, cold temperature, heat.Inflammatory disorders and related conditions which may be treated orprevented by use of the compositions of this invention include, but arenot limited to the following: arthritis, bronchitis, contact dermatitis,atophic dermatitis, psoriasis, seborrheic dermatitis, eczema, allergicdermatitis, polymorphous light eruptions, inflammatory dermatoses,folliculitis, alopecia, poison ivy, insect bites, acne inflammation,irritation induced by extrinsic factors including, but not limited to,chemicals, trauma, pollutants (such as cigarette smoke) and sunexposure, secondary conditions resulting from inflammation including butnot limited to xerosis, hyperkeratosis, pruritus, postinflammatoryhyperpigmentation, scarring and the like. Preferably, the inflammatorydisorders and related conditions which may be treated or prevented usingthe methods of the invention are arthritis, inflammatory dermatoses,contact dermatitis, allergic dermatitis, atopic dermatitis, polymorphouslight eruptions, irritation, including erythema induced by extrinsicfactors, acne inflammation, psoriasis, seborrheic dermatitis, eczema,poison ivy, insect bites, folliculitus, alopecia, and secondaryconditions and the like.

As used herein, unless otherwise specified, all percentages ofingredients in compositions are weight percent of active/solidsingredient based on the total weight of composition.

As used herein, a composition that is “essentially free” of aningredient means the composition that has about 2% or less of thatingredient by weight based on the total weight of the composition.Preferably, a composition that is essentially free of an ingredient hasabout 1% or less, more preferably about 0.5% or less, more preferablyabout 0.1% or less, more preferably about 0.05 or less, more preferablyabout 0.01% or less by weight based on the total weight of compositionof the ingredient. In certain more preferred embodiments, a compositionthat is essentially free of an ingredient is free of the ingredient,i.e. has none of that ingredient in the composition.

As used herein, “cosmetically/dermatologically acceptable” means thatthe ingredients which the term describes are suitable for use in contactwith tissues (e.g., the skin or hair) without undue toxicity,incompatibility, instability, irritation, allergic response, and thelike.

Any suitable extracts of Paulownia tomentosa wood may be used in accordwith the present invention. In general, the wood of the Paulowniatomentosa tree includes wood from the stem, branches, or a combinationof both. Suitable extracts of Paulownia tomentosa wood may be derivedfrom wood chips, wood dusts and/or small cuttings, and the like.

Suitable extracts of Paulownia tomentosa wood may be obtained usingconventional methods including, but not limited to, direct extraction ofmaterial from the wood by grinding, macerating, pressing, squeezing,mashing, centrifuging, and/or processes such as cold percolation,agitation/distillation, microwave assisted extraction, sonication,supercritical/subcritical CO₂ compressed gas extraction with or withoutpolar modifiers, pressurized solvent extraction, accelerated solventextraction, pressurized or normal hot water extraction, surfactantassisted pressurized hot water extraction, oil extraction, membraneextraction, Soxhlet extraction, the gold finger distillation/extractionand/or processes disclosed, for example, in U.S. Pat. Nos. 7,442,391,7,473,435, and 7537791 to Integrated Botanical Technologies, LLC,incorporated herein by reference, and the like, or by other methods suchas solvent extraction, and the like. Any of a variety of solventsincluding polar solvents, non-polar solvents, or combinations of two ormore thereof may be used in methods of comprising solvent extraction.Suitable polar solvents include polar inorganic solvents such as waterand the like, polar organic solvents such as alcohols and correspondingorganic acids, for example C₁-C₈ alcohols including methanol, ethanol,propanol, butanol, and the like and organic acids, including aceticacid, formic acid, propanoic acid, and the like, polyols and glycols,including C₁-C₈ polyols/glycols and the like, and combinations of two ormore thereof. Suitable non-polar solvents include non-polar organicsolvents such as alkanes, including C₁-C₈ alkanes, cycloalkanes,including C₁-C₈ alkanes, alkyl ethers, including C₁-C₈ alkyl ethers,Petroleum ethers, ketones, including C₁-C₈ ketones, methylene chloride,ethyl acetate, xylene, toluene, chloroform, vegetable oil, mineral oiland the like. In another embodiment extraction may be obtained bynon-polar solvents described above or supercritical fluid extractionwith or without a polar modifier such as C₁-C₈ alcohols, water, C₁-C₈polyols/glycols or C₁-C₈ organic acids. In certain preferredembodiments, the extract of the invention is a polar extract prepared bypulverizing the wood and extracting using a polar solvent having adielectric constant value of between 1 and 100 at 20° C., preferably adielectric constant of a value between 4 and 60 at 20° C., morepreferably a dielectric constant of a value between 4 and 50 at 20° C.,and even more preferably a dielectric constant of a value between 4 and40 at 20° C. Examples of preferred polar solvents include C₁-C₈alcohols, C₁-C₈ polyols/glycols, C₁-C₈ organic acids, water andcombinations of two or more thereof having a dielectric constant valueof between 1 and 100, preferably between 4 and 60, and more preferablybetween 5 and 40 at 20° C., including, but not limited to, thosesolvents and combinations of solvents having the desired dielectricconstant value as disclosed in “Dielectric Constants of Some OrganicSolvent-Water Mixtures at Various Temperatures,” Akerlof, Gosta; JACS,Vol. 54, No. 11 (November 1932), pp. 4125-4139, incorporated herein byreference. In certain preferred embodiments, the polar extract isextracted using one or more C₁-C₈ alcohols, C₁-C₈ polyols, C₁-C₈glycols, and combinations of two or more thereof. In certain morepreferred embodiments, the extract is extracted using one or more C₁-C₄alcohols, C₁-C₄ polyols, and/or C₁-C₄ glycols. In certain more preferredembodiments, the extract is prepared using a solvent comprisingmethanol, ethanol, or a combination thereof with or without presence ofwater. In more preferred embodiment, the extract is prepared usinganhydrous alcohol or reagent grade denatured alcohol and dried Kiri wooddust agitating at room temperature for 3 days. In certain preferredembodiments, the extract may be further refined by charcoal (alsoreferred to as active carbon) treatment.

In certain embodiments, the Paulownia tomentosa extract may be preparedto be essentially free of certain materials. In one embodiment, theextract is essentially free of Ursolic acid, beta-Sitosterol, or both.

In certain embodiments, the composition may additionally includeextracts from other parts of Paulownia tomentosa, for example, one ormore of the bark, leaves, roots, fruits, seeds, or flowers. In otherembodiments, the composition is essentially free from extracts of othernon-wood parts of Paulownia tomentosa.

Any suitable amounts of Paulownia tomentosa wood extract may be used inthe compositions of the present invention. Preferably, the compositionscomprise a safe and effective amount of Paulownia tomentosa woodextract. As used herein, a “safe and effective amount” means an amountof the extract or of the composition sufficient to induce the desiredeffect, but low enough to avoid serious side effects, includingcytotoxicity and the like. For embodiments comprising skin lighteninguses of the composition, a “skin lightening effective amount” means anamount of extract that is effective to achieve a ΔL value that isgreater than zero in the Skin Epidermal Equivalents Model as a skinLightening Test (ΔL) as described below. In certain preferredembodiments, the skin lightening effective amount is an amount effectiveto achieve a ΔL value of about 1 or greater.

For embodiments of the present invention related to uses of thecompositions for improving a sign of aging in skin, an “effective amountfor improving a sign of aging” means an amount that provides a percentinhibition of MMP-1 or MMP-9 production, measured in accord with theInhibition of UV-Induced MMP induction procedure of Example 11 below,that is greater than zero. In certain preferred embodiments, theeffective amount for improving a sign of aging is an amount thatprovides a percent inhibition of MMP-1 or MMP-9 production, measured inaccord with the Inhibition of UV-Induced MMP induction procedure ofExample 11 below, that is about 10% or greater.

For embodiments of the present invention related to uses of thecompositions for reducing inflammation, an “effective amount forreducing inflammation” means an amount that provides a percentinhibition of skin inflammation (IL-8), measured in accord with theAnti-Inflammatory effects on Release of UV-Induced Pro-inflammatorymediators on Reconstituted Epidermis procedure for IL-8 of Example 7below, that is greater than zero. In certain preferred embodiments, theeffective amount for improving a sign of aging is an amount thatprovides a percent inhibition of skin inflammation (IL-8), measured inaccord with the Anti-Inflammatory effects on Release of UV-InducedPro-inflammatory mediators on Reconstituted Epidermis procedure for IL-8of Example 7 below, that is about 10% or greater.

In certain preferred embodiments, the compositions comprise from greaterthan zero to about 20% Paulownia tomentosa wood extract. In certainother preferred embodiments, the compositions comprise from about 0.0001to about 20%, from about 0.001 to about 10%, from about 0.01 to about5%, from about 0.1 to about 5%, or from about 0.2 to about 2% Paulowniatomentosa wood extract. In certain other preferred embodiments, thecompositions comprise from greater than zero to about 1%, from about0.0001 to about 1%, from about 0.001 to about 1%, or from about 0.01 toabout 1% Paulownia tomentosa wood extract. In certain other preferredembodiments, the compositions comprise from about 1 to about 5%,preferably from about 2 to about 5% Paulownia tomentosa wood extract.

Any suitable carrier may be used in the compositions of the presentinvention. Preferably, for a skin care composition, the carrier is acosmetically-acceptable carrier. As will be recognized by those of skillin the art, cosmetically-acceptable carriers comprise carriers that aresuitable for use in contact with the body, in particular the skin forskin whitening applications, without undue toxicity, incompatibility,instability, irritation, allergic response, and the like. A safe andeffective amount of carrier is from about 50% to about 99.999%,preferably from about 80% to about 99.9%, more preferably from about99.9% to about 95%, most preferably from about 99.8% to about 98% of thecomposition. The carrier can be in a wide variety of forms. For example,emulsion carriers, including, but not limited to, oil-in-water,water-in-oil, water-in-oil-in-water, and oil-in-water-in-siliconeemulsions, are useful herein. These emulsions can cover a broad range ofviscosities, e.g, from about 100 cps to about 200,000 cps. Examples ofsuitable cosmetically-acceptable carriers includecosmetically-acceptable solvents and materials for cosmetic solutions,suspensions, lotions, creams, serums, essences, gels, toners, sticks,sprays, ointments, liquid washes and soap bars, shampoos, hairconditioners, pastes, foams, mousses, powders, shaving creams, wipes,patches, strips, powered patches, microneedle patches, bandages,hydrogels, film-forming products, facial and skin masks, make-up, liquiddrops, and the like. These product types may contain several types ofcosmetically-acceptable carriers including, but not limited tosolutions, suspensions, emulsions such as microemulsions andnanoemulsions, gels, solids, liposomes, other encapsulation technologiesand the like. The following are non-limitative examples of suchcarriers. Other carriers can be formulated by those of ordinary skill inthe art.

In one embodiment, the carrier contains water. In a further embodiment,the carrier may also contain one or more aqueous or organic solvents.Examples of organic solvents include, but are not limited to: dimethylisosorbide; isopropylmyristate; surfactants of cationic, anionic andnonionic nature; vegetable oils; mineral oils; waxes; gums; syntheticand natural gelling agents; alkanols; glycols; and polyols. Examples ofglycols include, but are not limited to, glycerin, propylene glycol,butylene glycol, pentalene glycol, hexylene glycol, polyethylene glycol,polypropylene glycol, diethylene glycol, triethylene glycol, caprylglycol, glycerol, butanediol and hexanetriol, and copolymers or mixturesthereof. Examples of alkanols include, but are not limited to, thosehaving from about 2 carbon atoms to about 12 carbon atoms (e.g., fromabout 2 carbon atoms to about 4 carbon atoms), such as isopropanol andethanol. Examples of polyols include, but are not limited to, thosehaving from about 2 carbon atoms to about 15 carbon atoms (e.g., fromabout 2 carbon atoms to about 10 carbon atoms) such as propylene glycol.The organic solvents may be present in the carrier in an amount, basedupon the total weight of the carrier, of from about 1 percent to about99.99 percent (e.g., from about 20 percent to about 50 percent). Watermay be present in the carrier (prior to use) in an amount, based uponthe total weight of the carrier, of from about 5 percent to about 95percent (e.g., from about 50 percent to about 90 percent). Solutions maycontain any suitable amounts of solvent, including from about 40 toabout 99.99%. Certain preferred solutions contain from about 50 to about99.9%, from about 60 to about 99%, from about 70 to about 99%, fromabout 80 to about 99%, or from about 90 to 99%.

A lotion can be made from such a solution. Lotions typically contain atleast one emollient in addition to a solvent. Lotions may comprise fromabout 1% to about 20% (e.g., from about 5% to about 10%) of anemollient(s) and from about 50% to about 90% (e.g., from about 60% toabout 80%) of water.

Another type of product that may be formulated from a solution is acream. A cream typically contains from about 5% to about 50% (e.g., fromabout 10% to about 20%) of an emollient(s) and from about 45% to about85% (e.g., from about 50% to about 75%) of water.

Yet another type of product that may be formulated from a solution is anointment. An ointment may contain a simple base of animal, vegetable, orsynthetic oils or semi-solid hydrocarbons. An ointment may contain fromabout 2% to about 10% of an emollient(s) plus from about 0.1% to about2% of a thickening agent(s).

The compositions useful in the present invention can also be formulatedas emulsions. If the carrier is an emulsion, from about 1% to about 10%(e.g., from about 2% to about 5%) of the carrier contains anemulsifier(s). Emulsifiers may be nonionic, anionic or cationic.

Lotions and creams can be formulated as emulsions. Typically suchlotions contain from 0.5% to about 5% of an emulsifier(s), while suchcreams would typically contain from about 1% to about 20% (e.g., fromabout 5% to about 10%) of an emollient(s); from about 20% to about 80%(e.g., from 30% to about 70%) of water; and from about 1% to about 10%(e.g., from about 2% to about 5%) of an emulsifier(s).

Single emulsion skin care preparations, such as lotions and creams, ofthe oil-in-water type and water-in-oil type are well-known in the artand are useful in the subject invention. Multiphase emulsioncompositions, such as the water-in-oil-in-water type or theoil-in-water-in-oil type, are also useful in the subject invention. Ingeneral, such single or multiphase emulsions contain water, emollients,and emulsifiers as essential ingredients.

The compositions of this invention can also be formulated as a gel(e.g., an aqueous, alcohol, alcohol/water, or oil gel using a suitablegelling agent(s)). Suitable gelling agents for aqueous and/or alcoholicgels include, but are not limited to, natural gums, acrylic acid andacrylate polymers and copolymers, and cellulose derivatives (e.g.,hydroxymethyl cellulose and hydroxypropyl cellulose). Suitable gellingagents for oils (such as mineral oil) include, but are not limited to,hydrogenated butylene/ethylene/styrene copolymer and hydrogenatedethylene/propylene/styrene copolymer. Such gels typically containsbetween about 0.1% and 5%, by weight, of such gelling agents.

The compositions of the present invention can also be formulated into asolid formulation (e.g., a wax-based stick, soap bar composition,powder, or wipe). The composition of the present invention can also becombined with a solid, semi-solid or dissolvable substrate (eg., a wipe,mask, pad, glove or strip).

The compositions of the present invention can also be formulated intoformulation used for the oral cavity, such as toothpaste, gel, rinse,solution, patch, and the like. The compositions may also be formulatedfor use in the eye, such as in solutions, emulsions, suspensions used asdrops or washes and the like, or formulated for use in the vaginalmucosa such as via gels, lotions, lubricants, and the like.

The compositions of the present invention may further comprise any of avariety of additional cosmetically active agents. Examples of suitableadditional active agents include: additional skin lightening agents,darkening agents, additional anti-aging agents, tropoelastin promoters,collagen promoters, anti-acne agents, shine control agents,anti-microbial agents such as anti-yeast agents, anti-fungal, andanti-bacterial agents, anti-inflammatory agents, anti-parasite agents,external analgesics, sunscreens, photoprotectors, antioxidants,keratolytic agents, detergents/surfactants, moisturizers, nutrients,vitamins, energy enhancers, anti-perspiration agents, astringents,deodorants, hair removers, hair growth enhancing agents, hair growthdelaying agents, firming agents, hydration boosters, efficacy boosters,anti-callous agents, agents for skin conditioning, anti-celluliteagents, fluorides, teeth whitening agents, anti-plaque agents, andplaque-dissolving agents, odor-control agents such as odor masking orpH-changing agents, and the like. Examples of various suitableadditional cosmetically acceptable actives include hydroxy acids,benzoyl peroxide, D-panthenol, UV filters such as but not limited toavobenzone (Parsol 1789), bisdisulizole disodium (Neo Heliopan AP),diethylamino hydroxybenzoyl hexyl benzoate (Uvinul A Plus), ecamsule(Mexoryl SX), methyl anthranilate, 4-aminobenzoic acid (PABA), cinoxate,ethylhexyl triazone (Uvinul T 150), homosalate, 4-methylbenzylidenecamphor (Parsol 5000), octyl methoxycinnamate (Octinoxate), octylsalicylate (Octisalate), padimate O (Escalol 507), phenylbenzimidazolesulfonic acid (Ensulizole), polysilicone-15 (Parsol SLX), trolaminesalicylate, Bemotrizinol (Tinosorb S), benzophenones 1-12, dioxybenzone,drometrizole trisiloxane (Mexoryl XL), iscotrizinol (Uvasorb HEB),octocrylene, oxybenzone (Eusolex 4360), sulisobenzone, bisoctrizole(Tinosorb M), titanium dioxide, zinc oxide, carotenoids, free radicalscavengers, spin traps, retinoids and retinoid precursors such asretinol, retinoic acid and retinol palmitate, ceramides, polyunsaturatedfatty acids, essential fatty acids, enzymes, enzyme inhibitors,minerals, hormones such as estrogens, steroids such as hydrocortisone,2-dimethylaminoethanol, copper salts such as copper chloride, peptidescontaining copper such as Cu:Gly-His-Lys, coenzyme Q10, amino acids sucha proline, vitamins, lactobionic acid, acetyl-coenzyme A, niacin,riboflavin, thiamin, ribose, electron transporters such as NADH andFADH2, and other botanical extracts such as oat, aloe vera, Feverfew,Soy, Shiitake mushroom extracts, and derivatives and mixtures thereof.

In certain preferred embodiments, the compositions of the presentinvention are skin care compositions that comprise an extract ofPaulownia tomentosa wood and at least one additional skin lighteningactive agent. Examples of suitable additional skin lightening activeagents include, but are not limited to, tyrosinase inhibitors,melanin-degradation agents, melanosome transfer inhibiting agentsincluding PAR-2 antagonists, exfoliants, sunscreens, retinoids,antioxidants, Tranexamic acid, tranexamic acid cetyl esterhydrochloride, skin bleaching agents, linoleic acid, adenosinemonophosphate disodium salt, Chamomilla extract, allantoin, opacifiers,talcs and silicas, zinc salts, and the like, and other agents asdescribed in Solano et al. Pigment Cell Res. 19 (550-571) and Ando etal. Int J Mol Sci 11 (2566-2575).

Examples of suitable tyrosinase inhibitors include but, are not limitedto, Vitamin C and its derivatives, Vitamin E and its derivatives, KojicAcid, Arbutin, resorcinols, hydroquinone, Flavones e.g. Licoriceflavanoids, Licorice root extract, Mulberry root extract, DioscoreaCoposita root extract, Saxifraga extract and the like, Ellagic acid,Salicylates and derivatives, Glucosamine and derivatives, Fullerene,Hinokitiol, Dioic acid, Acetyl glucosamine,5,5′-dipropyl-biphenyl-2,2′-diol (Magnolignan),4-(4-hydroxyphenyl)-2-butanol (4-HPB), combinations of two or morethereof, and the like. Examples of vitamin C derivatives include, butare not limited to, ascorbic acid and salts, Ascorbic Acid-2-Glucoside,sodium ascorbyl phosphate, magnesium ascorbyl phosphate, and naturalextract enriched in vitamin C. Examples of vitamin E derivativesinclude, but are not limited to, alpha-tocopherol, beta, tocopherol,gamma-tocopherol, delta-tocopherol, alpha-tocotrienol, beta-tocotrienol,gamma-tocotrienol, delta-tocotrienol and mixtures thereof, tocopherolacetate, tocopherol phosphate and natural extracts enriched in vitamin Ederivatives. Examples of resorcinol derivatives include, but are notlimited to, resorcinol, 4-substituted resorcinols like4-alkylresorcinols such as 4-butyresorcinol (rucinol), 4-hexylresorcinol(Synovea HR, Sytheon), phenylethyl resorcinol (Symwhite, Symrise),1-(2,4-dihydroxyphenyl)-3-(2,4-dimethoxy-3-methylphenyl)-Propane(nivitol, Unigen) and the like and natural extracts enriched inresorcinols. Examples of salicylates include, but are not limited to,4-methoxy potassium salicylate, salicylic acid, acetylsalicylic acid,4-methoxysalicylic acid and their salts. In certain preferredembodiments, the tyrosinase inhibitors include a 4-substitutedresorcinol, a vitamin C derivative, or a vitamin E derivative. In morepreferred embodiments, the tyrosinase inhibitor comprises Phenylethylresorcinol, 4-hexyl resorcinol, or ascorbyl-2-glucoside.

Examples of suitable melanin-degradation agents include, but are notlimited to, peroxides and enzymes such as peroxidases and ligninases. Incertain preferred embodiments, the melanin-inhibiting agents include aperoxide or a ligninase.

Examples of suitable melanosome transfer inhibiting agents includingPAR-2 antagonists such as soy trypsin inhibitor or Bowman-BirkInhibitor, Vitamin B3 and derivatives such as Niacinamide, Essentialsoy, Whole Soy, Soy extract. In certain preferred embodiments, themelanosome transfer inhibiting agents includes a soy extract orniacinamide.

Examples of exfolliants include, but are not limited to, alpha-hydroxyacids such as lactic acid, glycolic acid, malic acid, tartaric acid,citric acid, or any combination of any of the foregoing, beta-hydroxyacids such as salicylic acid, polyhydroxy acids such as lactobionic acidand gluconic acid, and mechanical exfoliation such as microdermabrasion.In certain preferred embodiments, the exfolliant include glycolic acidor salicylic acid.

Examples of sunscreens include, but are not limited to, avobenzone(Parsol 1789), bisdisulizole disodium (Neo Heliopan AP), diethylaminohydroxybenzoyl hexyl benzoate (Uvinul A Plus), ecamsule (Mexoryl SX),methyl anthranilate, 4-aminobenzoic acid (PABA), cinoxate, ethylhexyltriazone (Uvinul T 150), homosalate, 4-methylbenzylidene camphor (Parsol5000), octyl methoxycinnamate (Octinoxate), octyl salicylate(Octisalate), padimate 0 (Escalol 507), phenylbenzimidazole sulfonicacid (Ensulizole), polysilicone-15 (Parsol SLX), trolamine salicylate,Bemotrizinol (Tinosorb S), benzophenones 1-12, dioxybenzone,drometrizole trisiloxane (Mexoryl XL), iscotrizinol (Uvasorb HEB),octocrylene, oxybenzone (Eusolex 4360), sulisobenzone, bisoctrizole(Tinosorb M), titanium dioxide, zinc oxide, and the like.

Examples of retinoids include, but are not limited to, retinol (VitaminA alcohol), retinal (Vitamin A aldehyde), retinyl acetate, retinylpropionate, retinyl linoleate, retinoic acid, retinyl palmitate,isotretinoin, tazarotene, bexarotene, Adapalene, combinations of two ormore thereof and the like. In certain preferred embodiments, theretinoid is selected from the group consisting of retinol, retinal,retinyl acetate, retinyl propionate, retinyl linoleate, and combinationsof two or more thereof. In certain more preferred embodiments, theretinoid is retinol.

Examples of antioxidants include, but are not limited to, water-solubleantioxidants such as sulfhydryl compounds and their derivatives (e.g.,sodium metabisulfite and N-acetyl-cysteine, glutathione), lipoic acidand dihydrolipoic acid, stilbenoids such as resveratrol and derivatives,lactoferrin, iron and copper chelators and ascorbic acid and ascorbicacid derivatives (e.g., ascobyl-2-glucoside, ascorbyl palmitate andascorbyl polypeptide). Oil-soluble antioxidants suitable for use in thecompositions of this invention include, but are not limited to,butylated hydroxytoluene, retinoids (e.g., retinol and retinylpalmitate), tocopherols (e.g., tocopherol acetate), tocotrienols, andubiquinones. Natural extracts containing antioxidants suitable for usein the compositions of this invention, include, but not limited to,extracts containing flavonoids and isoflavonoids and their derivatives(e.g., genistein and diadzein), extracts containing resveratrol and thelike. Examples of such natural extracts include grape seed, green tea,black tea, white tea, pine bark, feverfew, parthenolide-free feverfew,oat extracts, blackberry extract, cotinus extract, soy extract, pomeloextract, wheat germ extract, Hesperedin, Grape extract, Portulacaextract, Licochalcone, chalcone, 2,2′-dihydroxy chalcone, Primulaextract, propolis, and the like.

The additional cosmetically active agent may be present in a compositionin any suitable amount, for example, in an amount of from about 0.0001%to about 20% by weight of the composition, e.g., about 0.001% to about10% such as about 0.01% to about 5%. In certain preferred embodiments,in an amount of 0.1% to 5% and in other preferred embodiments from 1% to2%.

In certain preferred embodiments, the compositions of the presentinvention are skin care compositions that comprise an extract ofPaulownia tomentosa wood and at least one additional anti-inflammatoryagent. Suitable additional anti-inflammatory active agents include, butare not limited to, compounds that have an IC50 (concentration at whicha compound achieves 50% inhibition of inflammation) of less than orequal to 100 μg/ml for Interleukin-2 in the ANTI-INFLAMMATORY ASSAY setforth below. In a preferred embodiment, the IC50 for the secondanti-inflammatory compounds is less than about 70 μg/ml, more preferablyless than about 50 μg/ml, more preferably less than about 40 μg/ml, morepreferably less than about 30 μg/ml.

The ANTI-INFLAMMATORY ASSAY assesses the ability of an agent to reducethe production of cytokines by human lymphocytes stimulated with theT-cell receptor (TCR) activating agent phytohaemagglutinin (PHA), and isconducted in the following manner. Human leukocytes are collected from ahealthy adult male via leukopheresis, and adjusted to a density of 1×10⁶cells/mL in serum free lymphocyte growth medium (ExVivo-15,Biowhittaker, Walkersville, Md.). PBLs are stimulated with 10 μg/mL PHAin the presence or absence of test samples following published methods(Hamamoto Y., et al. Exp Dermatol 2:231-235, 1993). Following a 48 hourincubation at 37° C. with 5% CO₂, the supernatant is removed andevaluated for cytokine content using commercially available multiplexcytokine detection kit.

Examples of suitable anti-inflammatory agents include substitutedresorcinols, (E)-3-(4-methylphenylsulfonyl)-2-propenenitrile (such as“Bay 11-7082,” commercially available from Sigma-Aldrich of St. Louis,Mo.), tetrahydrocurcuminoids (such as Tetrahydrocurcuminoid CG,available from Sabinsa Corporation of Piscataway, N.J.), extracts andmaterials derived from the following:

Phellodendron Amurense Cortex Extract (PCE)

Non-Denatured Soy (Glycine max)

Feverfew (Tanacetum parthenium)

Ginger (Zingiber officinale)

Ginko (Ginko Biloba)

Madecassoside (centella asiatica extract ingredient)

Cotinus (Cotinus coggygria)

Butterbur Extract (Petasites hybridus)

Goji Berry (Lycium barbarum)

Milk Thistle Extract (Silybum marianum))

Honeysuckle (Lonicera japonica)

Basalm of Peru (Myroxylon pereirae)

Sage (Salvia officinalis)

Cranberry Extract (Vaccinium oxycoccos)

Amaranth Oil (Amaranthus cruentus)

Pomegranate (Punica granatum)

Yerbe Mate (Ilex paraguariensis Leaf Extract)

White Lily Flower Extract (Lilium Candidum)

Olive Leaf Extract (Olea europaea)

Phloretin (apple extract)

Oat Flour (Aveena Sativa)

Lifenol (Hops: Humulus lupulus) Extract

Bugrane P (Ononis spinosa)

Licochalcone (Licorice: Glycyrrhiza inflate extract ingredient)

Symrelief (Bisabolol and Ginger extract)

combinations of two or more thereof, and the like.

Resorcinol is a dihydroxy phenol compound (I.e., 1,3 dihydroxybenzene)having by the following structure:

As used herein, “substituted resorcinol” means resorcinol comprising atleast one substituent in the 2, 4, 5, or 6 position. Thus, thesubstituted resorcinol may have as few as one and as many as foursubstituents. Positions 1 and 3 of the substituted resorcinol comprise—OH groups, as shown above.

In embodiments wherein substituted resorcinol is used foranti-inflammation, it is highly preferred that all of the substituentsof the substituted resorcinol are free of phenyl (—C₆H₅ aromatic)moieties. In certain embodiments, all of the substituents are free ofaromatic moieties (with or without heteroatoms). In certain suchembodiments, it is preferred that all of the substituents of thesubstituted resorcinol are free of ketone functionalities (carbonylsbonded to two other carbon atoms). In certain other such embodiments,all of the substituents of the substituted resorcinol are free of bothphenyl functionalities and ketone functionalities. In certain other suchembodiments, the substituted resorcinol comprises at least onesubstituent comprising 5 to 11 carbon atoms, preferably 5 to 10 carbonatoms, more preferably 5 to 9 carbon atoms, most preferably 5 to 8carbon atoms. In certain other such embodiments, at least onesubstituent comprises an alkyl group, such as one having the number ofcarbon atoms described above. The alkyl group is preferably unsaturated.

In certain embodiments, the 4 position of the resorcinol is substituted,and, in certain embodiments, only the 4 position is substituted. Inanother embodiment, the 4 position is substituted with an alkyl group.In certain preferred embodiments, the substituted resorcinol comprises asingle substituent at the 4 position that comprises an alkyl group. Incertain other preferred embodiments, the substituted resorcinolcomprises a single substituent at the 4 position that consists of analkyl group directly bonded to the benzene ring.

Particularly suitable substituted resorcinols for anti-inflammationagents include 4-hexyl resorcinol and 4-octylresorcinol, particularly4-hexyl resorcinol. The structures of 4-hexylresorcinol and4-octylresorcinol are shown below:

4-Hexyl resorcinol is commercially available as “SYNOVEA HR” fromSytheon of Lincoln Park, N.J. 4-Octylresorcinol is commerciallyavailable from City Chemical LLC of West Haven, Conn.

In certain embodiments, the substituted resorcinol comprises at leasttwo substituents in the 2, 4, 5, or 6 positions. Such substituents mayoptionally be linked to form a ring, such as a cyclic aliphatichydrocarbon optionally comprising heteroatoms such as sulfur or oxygen.Such a linked substituent may comprise 5 to 10 carbon atoms, e.g., 8 to10 carbon atoms, and optionally include 1 to 3 heteroatoms. Examples ofsuitable substituted resorcinols comprising cyclic aliphaticsubstituents joining the 2 and 3 positions include Zearalanone andβ-Zearalanol:

Zearalanone and β-Zearalanol are commercially available from SigmaChemicals of St. Louis, Mo.

In certain other embodiments, the substituted resorcinol compriseshalide-containing and/or nitroso-containing substituents. Suitableexamples contain —Cl or —N═O bonded directly to the benzene ring. Thesesubstituents may exist for example in the 2 and 4, 2 and 6, or 4 and 6positions. An example of a dihalide-substituted resorcinol is2,6-dichlororesorcinol. An example of a dinitroso-substituted resorcinolis 2,4-dinitrososorcinol:

2,6-Dichlororesorcinol and 2,4-Dinitrososorcinol are available from CityChemical LLC of West Haven, Conn.

Substituted resorcinols are prepared by means known in the art, forexample, using techniques described in U.S. Pat. No. 4,337,370, thecontents of which are incorporated herein by reference.

The substituted resorcinols may have any suitable molecular weight. Incertain embodiments, the molecular weight of the substituted resorcinolranges between about 175 and about 300.

By “extracts of feverfew,” it is meant extracts of the plant “Tanacetumparthenium,” such as may be produced according to the details set forthe in US Patent Application Publication No. 2007/0196523, entitled“PARTHENOLIDE FREE BIOACTIVE INGREDIENTS FROM FEVERFEW (TANACETUMPARTHENIUM) AND PROCESSES FOR THEIR PRODUCTION.” One particularlysuitable feverfew extract is commercially available as about 20% activefeverfew, from Integrated Botanical Technologies of Ossining, N.Y.

Compositions of the present invention may include a cosmeticallyeffective amount of one or more additional anti-inflammatory compounds.The compositions preferably include, on an active basis, from about 0.1%to about 10%, more preferably from about 0.5% to about 5%, of theadditional anti-inflammatory compound.

In the inventive composition, the ratio of the concentrations of thePaulownia tomentosa wood extract to the additional anti-inflammatorycompound may be varied. For example, the extract and theanti-inflammatory compound may be present in a concentration by weightratio (which is determined by dividing the concentration by weight ofthe dry extract by the concentration by weight of the additionalanti-inflammatory compound) of about 0.001 to about 100, preferablyabout 0.01 to about 10, more preferably about 0.25 to about 2.

In certain preferred embodiments, the compositions of the presentinvention are skin care compositions that comprise an extract ofPaulownia tomentosa wood and at least one additional agent improving thesigns of aging. Examples of suitable additional agents improving thesigns of aging include, but are not limited to, tropoelastin promoters,collagen promoters, retinoids, hyaluronic acid, dimethylaminoethanol,N,N,N′,N′-Tetrakis(2-hydroxypropyl)ethylenediamine, alpha hydrox acids,polyhydroxyacids, and combinations of two or more thereof.

“Tropoelastin promoter,” as used herein, refers to a class of compoundsthat possess the biological activity of enhancing the production oftropoelastin. Tropoelastin promoters, according to the presentinvention, include all natural or synthetic compounds that are capableof enhancing the production of tropoelastin in the human body.

Suitable tropoelastin promoters may be determined, for example, usingthe TROPOELASTIN PROMOTER ASSAY. The TROPOELASTIN PROMOTER ASSAY isperformed as follows. Rat cardiac myoblasts H9C2 (which may bepurchased, for example from ATCC of Manassas, Va.) are used. Culturesare maintained in Dulbecco's modified Eagle's medium (DMEM, InvitrogenLife Technologies, Carlsbad, Calif.) supplemented with 10% fetal bovineserum, 2 mM glutamine, 100 units/ml penicillin, and 50 ug/mlstreptomycin (Invitrogen LifeTechnologies, Carlsbad, Calif.). Cellcultures are transiently transfected with the elastinpromoter-luciferase reporter construct (Elp2.2, a 2.2 kb elastinpromoter fragment from nt −2267 to nt+2, driving the firefly luciferasegene, which may be obtained from Promega, Madison Wis.). DNA is preparedby Qiagen Maxi columns (Qiagen Valencia, Calif.). In all transfections,a construct with the thymidine kinase promoter and the Renillaluciferase reporter gene (pRL-TK, Promega, Madison Wis.) is included asan internal control. Typically, cells grown in 48-well plates aretransfected with 0.45 ug total DNA per well using Lipofectamine 2000(Invitrogen Life Technologies, Carlsbad, Calif.). One day aftertransfection, cells are treated with agents at indicated concentrationsfor approximately 24 hours before they are lysed for luciferase assays,using Dual-Luciferase Reporter System from Promega (Madison, Wis.),following manufacturer's protocol. The firefly luciferase activity ismeasured first (representing elastin promoter activity), followed by therenilla luciferase (internal control), using luminometer LMAX, fromMolecular Devices (Sunnyvale, Calif.). The ratio of these two luciferaseactivities (RLU) is used to evaluate the Tropoelastin Promoter Activity.The tropoelastin promoter preferably has a Tropoelastin PromoterActivity of at least 1.1, preferably at least 1.25, more preferably atleast 1.3, and most preferably at least 1.5, at least one concentrationin the range of 0.5 micrograms/milliliter to 2.5 milligrams permilliliter (on an actives basis), and preferably at least oneconcentration in the range of 1.0 micrograms/milliliter to 2.5milligrams per milliliter (on an actives basis).

Examples of suitable tropoelastin promoters include, but are not limitedto, blackberry extracts, cotinus extracts, feverfew extracts, extractsof Phyllanthus niruri and bimetal complexes having copper and/or zincconstituents. The bimetal complex having copper and/or zinc constituentsmay be, for example, copper-zinc citrate, copper-zinc oxalate,copper-zinc tartarate, copper-zinc malate, copper-zinc succinate,copper-zinc malonate, copper-zinc maleate, copper-zinc aspartate,copper-zinc glutamate, copper-zinc glutarate, copper-zinc fumarate,copper-zinc glucarate, copperzinc polyacrylic acid, copper-zinc adipate,copper-zinc pimelate, copper-zinc suberate, copper-zinc azealate,copper-zinc sebacate, copper-zinc dodecanoate, or combinations thereof.In a preferred embodiment, the tropoelastin promoter is selected fromblackberry extracts, cotinus extracts, feverfew extracts, andcombinations thereof. In a particularly preferred embodiment, thetropoelastin promoter is selected from blackberry extracts, feverfewextracts, and combinations thereof.

By “cotinus extract,” it is meant an extract of the leaves of “Cotinuscoggygria,” such as a water extract thereof, available from Bilkokoop ofSofia, Bulgaria.

By “blackberry extract,” it is meant a blend of compounds isolated fromthe plant of the genus Rubus, and preferably Rubus fruticosus. In oneembodiment, the compounds are isolated from the flowers of the plant. Ina further embodiment, the compounds are isolated from dried flowers ofthe plant. Such compounds may be isolated from one or more part of theplant (e.g., the whole plant, flower, seed, root, rhizome, stem, fruitand/or leaf of the plant). In a preferred embodiment, the blackberryextract is a blackberry leaf extract.

The extraction process may include by physically removing a piece ofsuch plant, and, for example, grinding it. Further extraction ofsuitable compounds may also be isolated from the plant by usingextraction procedures well known in the art (e.g., the use of organicsolvents such as lower C1-C8 alcohols, C1-C8 alkyl polyols, C1-C8 alkylketones, C1-C8 alkyl ethers, acetic acid C1-C8 alkyl esters, andchloroform, and/or inorganic solvents such as water, inorganic acidssuch as hydrochloric acid, and inorganic bases such as sodiumhydroxide).

For example, a blackberry leaf extract may be prepared by an extractionwith water, alcohols such as ethanol or combination thereof as thesolvent. However, an extract produced with a solvent including bothethanol and water is preferred. The blackberry leaves are preferablydried prior to extraction. It is also preferable to use only the leavesof the blackberry plant for the extraction and not also other plantparts such as the fruit (berries) of the blackberry or its branches androots. In one embodiment, the extraction process for the production of ablackberry leaf extract comprises the following steps: a) addition toblackberry leaves of an solvent containing an alcohol selected from thegroup consisting of methanol, ethanol, npropanol, isopropanol, b)Extraction of the blackberry leaves with the solvent for up to 72 hours.

Detailed procedures for preparing a suitable blackberry leaf extract aredisclosed in US Patent Application Publication No. 2008/0095719, thedisclosure of which is incorporated herein in its entirety.

One particularly suitable blackberry extract is produced by extractingthe leaves of Rubus fruticosus with a mixture of water and ethanolcompounded to an activity of about 5% to about 10%, with a maltodextrinmatrix, commercially available from Symrise Inc. of Teterboro, N.J., andis sold under the name “SymMatrix.”

Extracts of “Phyllanthus niruri” may be harvested and used as the wholeplant, or optionally one or more parts of the plant (e.g., flower, seed,root, rhizome, stem, fruit and/or leaf of the plant) may be used. ThePhyllanthus niruri plant or parts thereof may be finely divided, such asby grinding or milling, to a powder. A suitable milled form ofPhyllanthus niruri is commercially available from Raintree Nutrition,Inc., of Carson City, Nev. Preferably, a low molecular weight fractionof Phyllanthus niruri is used, for instance a fraction of Phyllanthusniruri substantially free of molecular species having a molecular weightof greater than about 100,000 daltons. Preferably, such low molecularweight fraction is water extractable from the Phyllanthus niruri plant.

Compositions of the present invention may include a cosmeticallyeffective amount of one or more tropoelastin promoters such as thosedescribed above. The compositions preferably include, on an activebasis, from about 0.1% to about 10% of the tropoelastin promoters, morepreferably from about 0.5% to about 5% of tropoelastin promoters, andmost preferably from about 0.5% to about 2% of the tropoelastinpromoters.

“Collagen promoter,” as used herein, refers to compounds that possessthe biological activity of enhancing the production of collagen.“Non-retinoid collagen promoters”, according to the present invention,include all natural or synthetic compounds that are not retinoids, orderived from retinoids, and are capable of enhancing the production ofcollagen in the human body.

Suitable collagen promoters may be determined, for example, using theCOLLAGEN PROMOTER ASSAY. The COLLAGEN PROMOTER ASSAY is performed asfollows. Rat cardiac myoblasts H9C2, which may be purchased from ATCC(Manassas, Va.), are used. Cultures are maintained in Dulbecco'smodified Eagle's medium (DMEM, Invitrogen Life Technologies, Carlsbad,Calif.) supplemented with 10% fetal bovine serum, 2 mM glutamine, 100units/ml penicillin, and 50 ug/ml streptomycin (Invitrogen lifetechnologies, Carlsbad, Calif.). Cell cultures are transientlytransfected with the Collagen1A promoter-luciferase reporter construct,driving the firefly luciferase gene, which may obtained for example fromPREMAS Biotech Pvt. Ltd (Haryana, India). In all transfections, aconstruct with the thymidine kinase promoter and the Renilla luciferasereporter gene (pRL-TK, Promega, Madison, Wis.) is included as aninternal control. Cells grown in 48-well plates are transfected with0.45 ug total DNA per well using Lipofectamine 2000 (Invitrogen lifetechnologies, Carlsbad, Calif.). One day after transfection, cells aretreated with agents at the indicated concentrations for approximately 24hours before they are lysed for luciferase assays, using Dual-LuciferaseReporter System from Promega (Madison, Wis.), following manufacturer'sprotocol. The firefly luciferase activity is measured first(representing collagen promoter activity), followed by the renillaluciferase (internal control), using luminometer LMAX, from MolecularDevices (Sunnyvale, Calif.). The ratio of these two luciferaseactivities (RLU) is used to evaluate the activity of each promoter.

The suitable collagen promoter preferably has a Collagen PromoterActivity of at least 1.2, preferably at least 1.25, more preferably atleast 1.3; at least one concentration in the range of 0.5micrograms/milliliter to 2.5 milligrams per milliliter (on an activesbasis), preferably at least one concentration in the range of 1.0micrograms/milliliter to 2.5 milligrams per milliliter (on an activesbasis).

Examples of suitable non-retinoid collagen promoters include, but arenot limited to the following: extracts of feverfew (Tanacetumparthenium), extracts of Centella asiatica, extracts of Siegesbeckiaorientalis; extracts of soy; collagenpromoting peptides; ursolic acid;and asiaticoside.

Centella asiatica, also known as Violette marronne on Reunion Island,Gotu Kola or Indian pennywort in India, Centella repanda in NorthAmerica, and Talapetraka in Madagascar, is a polymorphous herb andbelongs to the family of Umbelliferae (Apiaceae), particularly to theHydrocotyle subfamily. It grows wild throughout the tropics and prefersmoist and shady regions at an altitude of about 600 to 1200 meters abovesea level. Centella asiatica has three varieties: Typica, Abyssinica,and Floridana. The herb is known and used for its healing, sedative,analgesic, antidepressant, antiviral and antimicrobial properties. Thebiological activity of the herb appears to be due to the presence oftriterpene molecules in the herb. A suitable extract of Centellaasiatica is available as TECA from Bayer Consumer HealthCare of Basel,Switzerland.

By “extracts of Siegesbeckia orientalis,” is meant any of variousextracts of the plant Siegesbeckia orientalis, including Darutosideavailable from Sederma (Croda International Group of Edison, N.J.).

Suitable collagen-promoting peptides include the following:

(1) matrikine peptides, (i.e., a peptide derived from the degradation ofextracellular matrix proteins-collagen, elastin, or proteoglycan)including palmitoyl pentapeptides, in particularPal-Lys-Thr-Thr-Lys-Ser-OH, available as MATRIXYL from Sederma (CrodaInternational Group of Edison, N.J.);

(2) GHK copper peptide available as PROCYTE from Photomedex ofMontgomeryville, Pa.;

(3) Palmitoyl GHK peptide available as Biopoeptide CL from Sederma(Croda International Group of Edison, N.J.);

(4) Peptides VFTRN, TRNDKL disclosed in EP1775306 B1, and describedbelow in the following formulas I, II and III:

wherein formula I contains at least six amino acid residues; and:A1 is Val, Ala, Leu, Met or absent;A2 is Arg, Lys or absent;A3 is Phe, Tyr or absent;A4 is Thr, Ser, Ala, or Lys;A5 is Arg or Lys;A6 is Asn, Asp, Gly, or Gln;A7 is Asp, Asn, Glu, or absent;A8 is Lys, Arg or absent; andA9 is Leu, Met, Val, Ile, Phe or absent;provided that A3 may only be absent if A2 is absent, A2 may only beabsent if A1 is absent, A7 may be absent only if A8 is absent, and A8may only be absent if A9 is absent;each R1 and R2, independently, is H, C1-12 alkyl, C7-10 phenylalkyl, orC(═O)E1, where E1 is C1-12 alkyl, C3-14 alkenyl, C3-14 alkynyl, phenyl,3,4-dihydroxyphenylalkyl, naphthyl, or C7-10 phenylalkyl; provided thatwhen either R1 or R2 is C(═O)E1, the other must be H; andR3 is OH, NH2, C1-12 alkoxy, C7-10 phenylalkoxy, C11-14 naphthylalkoxy,C1-12 alkylamino, C7-10 phenylalkylamino, or C11-14 naphthylalkylamino;or a cosmetically acceptable salt thereof

wherein formula II contains at least six amino acid residues; and:A′ 1 is Val, Ala, Leu or Met;A′2 is Arg or Lys;A′3 is Phe or Tyr;A′4 is Leu, Met, Val, Ile or Phe;A′5 is His, Tyr or Phe;A′6 is Ser, Thr, Ala or Lys;A′7 is Tyr or Phe;A′8 is Asp, Asn or Glu;A′9 is Leu, Met, Val, Ile or Phe;A′10 is Lys or Arg;A′11 is Asn, Asp, Gly or Gln; andR1, R2, and R3, are the same as those defined in formula I.

wherein formula III contains at least six amino acid residues; and:A″1 is Cys or Ser;A″2 is His, Tyr or Phe;A″3 is Lys or Arg;A″4 is Leu, Met, Val, Ile or Phe;A″5 is Leu, Met, Val, Ile or Phe;A″6 is His, Tyr or Phe;A″7 is Asn, Asp, Gly or Gln;A″8 is Val, Ala, Leu or Met;A″9 is Asn, Asp, Gly or Gln;A″10 is Lys or Arg; andR1, R2, and R3, are the same as those defined in formula I.

(5) Biomimetic tetrapeptides, such as those available as Chronoline TriPeptide from Unipex of Québec, Canada; and

(6) Palmitoyl tri-peptide, available as Syn-Coll from DSM of Basel,Switzerland. Ursolic acid is also known as pentacyclic triterpene acid,Prunol, Malol, Urson, beta-ursolic acid and3-Beta-Hydroxy-Urs-12-En-28-Oic Acid, It is commercially available forexample from Sigma-Aldrich of St. Louis, Mo.

Asiaticoside, also known chemically as:[6-[[3,4-dihydroxy-6-(hydroxymethyl)-5-(3,4,5-trihydroxy-6-methyloxan-2-yl)oxyoxan-2-yl]oxymethyl]-3,4,5-trihydroxyoxan-2-yl]10,11-dihydroxy-9-(hydroxymethyl)-1,2,6a,6b,9,12a-hexamethyl-2,3,4,5,6,6a,7,8,8a,10,11,12,13,14b-tetradecahydro-1H-picene-4-a-carboxylate)is commercially available for example from Bayer Sante FamilialeDivision Serdex, 69, Boulevard Victor Hugo 93400 SAINT-OUEN France.

Compositions of the present invention may include a cosmeticallyeffective amount of one or more collagen promoters. The compositionspreferably include, on an active basis, from about 0.1% to about 10% ofthe collagen promoters, more preferably from about 0.5% to about 5% ofcollagen promoters, and most preferably from about 0.5% to about 2% ofthe collagen promoters.

A variety of other materials may also be present in the compositions ofthe present invention. In certain preferred embodiments, the compositionis a skin care composition comprising one or more materials selectedfrom the group consisting of: surfactants, chelating agents, emollients,humectants, conditioners, preservatives, opacifiers, fragrances and thelike.

What is meant by an emollient is a compound that helps to maintain thesoft, smooth, and pliable appearance of the skin (e.g., by remaining onthe skin surface or in the stratum corneum to act as a lubricant).Examples of suitable emollients include those found in Chapter 35, pages399-415 (Skin Feel Agents, by G Zocchi) in Handbook of Cosmetic Scienceand Technology (edited by A. Barel, M. Paye and H. Maibach, Published in2001 by Marcel Dekker, Inc New York, N.Y.), and include, but are notlimited to, petrolatum, hexyldecyl stearate and plant, nut, andvegetable oils such as macadamia nut oil, rice bran oil, grape seed oil,palm oil, prim rose oil, hydrogenates peanut oil, and avocado oil.

What is meant by a humectant is a compound intended to increase thewater content of the top layers of skin (e.g., hygroscopic compounds).Examples of suitable humectants include those found Chapter 35, pages399-415 (Skin Feel Agents, by G Zocchi) in Handbook of Cosmetic Scienceand Technology (edited by A. Barel, M. Paye and H. Maibach, Published in2001 by Marcel Dekker, Inc New York, N.Y.) and include, but are notlimited to, glycerin, sorbitol or trehalose (e.g., α,α-trehalose,β,β-trehalose, α,β-trehalose) or a salt or ester thereof (e.g.,trehalose 6-phosphate).

What is meant by a surfactant is a surface-active agent intended tocleanse or emulsify. Examples of suitable surfactants include thosefound in Chapter 37, pages 431-450 (Classification of surfactants, by L.Oldenhove de Guertechin) in Handbook of Cosmetic Science and Technology(edited by A. Barel, M. Paye and H. Maibach, Published in 2001 by MarcelDekker, Inc New York, N.Y.) and include, but are not limited to anionicsurfactants such as sulfates, cationic surfactants such as betaines,amphoteric surfactants such as sodium coco glycinate, noionicsurfactants such as alkyl polygucosides.

Examples of suitable chelating agents include those which are capable ofprotecting and preserving the compositions of this invention.Preferably, the chelating agent is ethylenediamine tetracetic acid(“EDTA”), and more preferably is tetrasodium EDTA, availablecommercially from Dow Chemical Company of Midland, Mich. under thetradename, “Versene 100XL.”

Suitable preservatives include, for example, parabens, quaternaryammonium species, phenoxyethanol, benzoates, DMDM hydantoin, organicacids and are present in the composition in an amount, based upon thetotal weight of the composition, from about 0 to about 1 percent or fromabout 0.05 percent to about 0.5 percent.

Any of a variety of conditioners which impart additional attributes,such as gloss to the hair are suitable for use in this invention.Examples include, but are not limited to, volatile silicone conditioningagent having an atmospheric pressure boiling point less than about 220°C. Examples of suitable volatile silicones nonexclusively includepolydimethylsiloxane, polydimethylcyclosiloxane, hexamethyldisiloxane,cyclomethicone fluids such as polydimethylcyclosiloxane availablecommercially from Dow Corning Corporation of Midland, Mich. under thetradename, “DC-345” and mixtures thereof, and preferably includecyclomethicone fluids. Other suitable conditioners include cationicpolymers, including polyquarterniums, cationic guar, and the like.

Any of a variety of commercially available pearlescent or opacifyingagents are suitable for use in this invention. Examples of suitablepearlescent or opacifying agents include, but are not limited to, monoor diesters of (a) fatty acids having from about 16 to about 22 carbonatoms and (b) either ethylene or propylene glycol; mono or diesters of(a) fatty acids having from about 16 to about 22 carbon atoms (b) apolyalkylene glycol of the formula: HO-(JO)_(a)—H, wherein J is analkylene group having from about 2 to about 3 carbon atoms; and a is 2or 3; fatty alcohols containing from about 16 to about 22 carbon atoms;fatty esters of the formula: KCOOCH₂L, wherein K and L independentlycontain from about 15 to about 21 carbon atoms; inorganic solidsinsoluble in the shampoo composition, and mixtures thereof.

Any fragrance compositions suitable for use on skin and desirable for askin care composition may be used in accord with the present invention.

In certain preferred embodiments, the present invention is in the formof a substrate comprising a composition of the present invention. Anysuitable substrate may be used in the present invention. Examples ofsuitable substrates and substrate materials are disclosed, for example,in U.S. Published Application Nos. 2005/0226834 and 2009/0241242 whichare incorporated herein by reference in their entirety.

In certain preferred embodiments, the substrate is a wipe, glove, or afacial mask. Preferably, such embodiments comprise a water-insolublesubstrate as such is defined in the cited references above. For certainembodiments, the water-insoluble substrate may have a size and shapesuch that it covers the face of a human user to facilitate placing thewater-insoluble substrate about the face of the user as a masksubstrate. For example, the water-insoluble mask substrate may haveopenings for a mouth, nose, and/or eyes of the user. Alternatively, thewater-insoluble substrate may have no such openings. Such aconfiguration without openings may be useful for embodiments of theinvention in which the water-insoluble substrate is intended to bedraped over a non-facial expanse of skin or if the water-insolublesubstrate is intended to be used as wipe. The water-insoluble substratemay have various shapes, such as an angular shape (e.g., rectangular) oran arcuate shape such as circular or oval. For certain embodiments, thesubstrate is a glove such as described in U.S. Published Application No2006/0141014 which is incorporated herein in its entirety.

In one embodiment of the invention, the product includes a plurality ofwater-insoluble substrates of different shapes. In one embodiment of theinvention, the product includes a first water-insoluble substrate and asecond water-insoluble substrate. The first water-insoluble substrate isshaped for application onto the forehead and the second water-insolublesubstrate is shaped for application proximate to the mouth, such asareas above and/or below the lips, the chin, and/or the cheeks. In oneembodiment of the invention, the first water-insoluble substrate is alsoapplied to the nose region of the face. The first water-insolublesubstrate may have a surface area of from about 100 cm² to about 200cm², such as from about 120 cm² to about 160 cm² and the secondwater-insoluble substrate has a surface area of from about 100 cm² toabout 300 cm², such as from about 150 cm² to about 250 cm². In oneembodiment of the invention, the water-insoluble substrate has a lowstiffness such that it may, for example, readily drape over or conformto the face or other body parts of the user.

The present invention further comprises methods of lightening the skinby applying to skin in need of skin lightening treatment an extract ofPaulownia tomentosa wood, as such extracts and embodiments thereof aredescribed above. In certain embodiments, the method comprises applying acomposition of the present invention comprising an extract of Paulowniatomentosa wood, as such compositions are described above in variousembodiments, to skin in need of skin lightening treatment.

The present invention may comprise application to any skin in need oftreatment on the body. For example, application may be made to any oneor more of the skin of the face, lips, neck, chest, back, arms, axilla,hands, feet and/or legs.

Preferably, the methods of the present invention comprise applying askin lightening effective amount of Paulownia tomentosa wood extract tothe skin, preferably a safe and effective amount. In certain preferredembodiments, the methods comprise applying from greater than zero toabout 20% Paulownia tomentosa wood extract to the skin in need. Incertain other preferred embodiments, the methods comprise applying fromabout 0.0001 to about 20%, from about 0.001 to about 10%, from about0.01 to about 5%, from about 0.1 to about 5%, or from about 0.2 to about2% Paulownia tomentosa wood extract to the skin in need. In certainother preferred embodiments, the methods comprise from greater than zeroto about 1%, from about 0.0001 to about 1%, from about 0.001 to about1%, or from about 0.01 to about 1% Paulownia tomentosa wood extract tothe skin. In certain other preferred embodiments, the methods compriseapplying from about 1 to about 5%, preferably from about 2 to about 5%Paulownia tomentosa wood extract to the skin.

Any suitable method of applying the extract to the skin in need may beused in accord with the present invention. For example, the extract maybe applied directly from a package to the skin in need, by hand to theskin in need, or may be transferred from a substrate such as a wipe ormask, or a combination of two or more thereof. In other embodiments, theextract may be applied via a dropper, tube, roller, spray, patch oradded to a bath or otherwise to water to be applied to the skin, and thelike.

In certain embodiments, the methods of the present invention furthercomprise the step of leaving the Paulownia tomentosa wood extract incontact with the skin for period of time. For example, in certainpreferred embodiments after application, the extract is left in contactwith the skin for a period of about 15 minutes or greater. In certainmore preferred embodiments, the extract is left in contact with the skinfor about 20 minutes or greater, more preferably about 1 hour orgreater.

In certain embodiments, the method of the present invention comprises aregimen comprising applying the Paulownia tomentosa wood extract to skinmultiple times over a selected period of time. For example, in certainembodiments, the present invention provides a method of skin lighteningcomprising applying to skin in need of skin lightening a compositioncomprising a Paulownia tomentosa wood extract once or twice daily for atleast 12 weeks, preferably at least 8 weeks and more preferably for atleast 2 weeks.

In certain preferred embodiments, the methods of the present inventioncomprise applying at least two different compositions or productscomprising Paulownia tomentosa wood extract to the skin. For example,the methods may comprise applying a first composition comprisingPaulownia tomentosa wood extract to skin in need of skin lighteningfollowed by applying a second composition comprising Paulownia tomentosawood extract, but that is otherwise different from the firstcomposition, to the skin in need of skin lightening. In certainpreferred embodiments, the first and second composition may beindependently selected from the group consisting of lotions, cleansers,masks, wipes, creams, serums, gels, and the like. In certain preferredembodiments, at least one of the first and second compositions is acleanser, lotion, cream, essence, or serum and the other is a facialmask or wipe. In certain other preferred embodiments, at least one ofthe first and second compositions is a cleanser and the other is alotion or cream.

In certain other preferred embodiments, the method comprises applying atleast three products comprising Paulownia tomentosa wood extract to skinin need of skin lightening. Preferably such three products are selectedfrom the group consisting of cleansers, lotions, creams, essences, andfacial masks.

The present invention further comprises methods of improving a sign ofaging in skin comprising the step of applying to skin in need ofimproving the signs of aging an extract of Paulownia tomentosa wood assuch extracts and compositions thereof are described above, as well asmethods of reducing skin inflammation comprising the step of applying toa skin in need of reducing skin inflammation an extract of Paulowniatomentosa wood as such extracts and compositions thereof are describedabove.

The present invention may comprise application to any skin in need oftreatment on the body. For example, application may be made to any oneor more of the skin of the face, lips, neck, chest, back, arms, axilla,hands, feet and/or legs.

Preferably, the methods of the present invention comprise applying asafe and effective amount of Paulownia tomentosa wood extract to theskin. In certain preferred embodiments, the methods comprise applyingfrom greater than zero to about 20% Paulownia tomentosa wood extract tothe skin in need. In certain other preferred embodiments, the methodscomprise applying from about 0.0001 to about 20%, from about 0.001 toabout 10%, from about 0.01 to about 5%, from about 0.1 to about 5%, orfrom about 0.2 to about 2% Paulownia tomentosa wood extract to the skinin need. In certain other preferred embodiments, the methods comprisefrom greater than zero to about 1%, from about 0.0001 to about 1%, fromabout 0.001 to about 1%, or from about 0.01 to about 1% Paulowniatomentosa wood extract to the skin. In certain other preferredembodiments, the methods comprise applying from about 1 to about 5,preferably from about 2 to about 5% Paulownia tomentosa wood extract tothe skin.

Any suitable method of applying the extract to the skin in need may beused in accord with the present invention. For example, the extract maybe applied directly from a package to the skin in need, by hand to theskin in need, or may be transferred from a substrate such as a wipe ormask, or a combination of two or more thereof. In other embodiments, theextract may be applied via a dropper, tube, roller, spray, patch oradded to a bath or otherwise to water to be applied to the skin, and thelike.

In certain embodiments, the methods of the present invention furthercomprise the step of leaving the Paulownia tomentosa wood extract incontact with the skin for period of time. For example, in certainpreferred embodiments after application, the extract is left in contactwith the skin for a period of about 15 minutes or greater. In certainmore preferred embodiments, the extract is left in contact with the skinfor about 20 minutes or greater, more preferably about 1 hour orgreater.

In certain embodiments, the method of the present invention comprises aregimen comprising applying the Paulownia tomentosa wood extract to skinmultiple times over a selected period of time. For example, in certainembodiments, the present invention provides a method comprising applyingto skin in need a composition comprising a Paulownia tomentosa woodextract once or twice daily for at least 12 weeks, preferably at least 8weeks and more preferably for at least 2 weeks.

In certain preferred embodiments, the methods of the present inventioncomprise applying at least two different compositions or productscomprising Paulownia tomentosa wood extract to the skin. For example,the methods may comprise applying a first composition comprisingPaulownia tomentosa wood extract to skin in need followed by applying tosuch skin a second composition comprising Paulownia tomentosa woodextract, but that is otherwise different from the first composition, tothe skin in need.

The compositions of the present invention may be suitable for a varietyof other uses. For example, compositions of the present invention may beuseful for cleansing and/or moisturizing dry skin, treating signs ofaging and/or for treating inflammation, including post-inflammatoryhyperpigmentation, for reducing pore size, acne treatment, for reducingsebum production, for scar mitigation and reducing the appearance ofstretch marks, for reducing the appearance of cellulite or orange peelappearance. In certain other embodiments, compositions of the presentinvention may be applied simultaneously with or within several hours ofa mechanical or physical exfoliant such as a microdermabrasiontreatment, or with a chemical exfoliant or keratolytic agent such assalicylic acid. In certain other embodiments, compositions of thepresent invention are applied to mucosa or other tissue such as vaginal,oral, or ocular tissue. In certain other embodiments, compositions ofthe present invention are applied to mild wounds or post-surgical sitesto facilitate healing, to insect bites, to poison ivy or similar skinconditions, or in general to mitigate itch. In certain otherembodiments, compositions of the present invention are applied tomitigate skin irritations. The irritation may be of external originscaused by ingredients in skin care and cosmetic products such asretinoid and its derivatives, benzyol peroxide, alpha-hydroxy acids andderivatives thereof, salicylic acid, surfactants, natural plantextracts, sunscreen actives, urea, and preservatives, etc. Theirritation may be of other external origins such as the sun, wind, orshaving. The irritation may also be caused by inherent diseaseconditions such as acne, rosacea, atopic dermatitis, and other diseasestates. In other embodiments, compositions of the present invention maybe useful to reduce redness of the gums. The extracts may further besuitable for use in reducing the appearance of telangiectasia or spiderveins, reducing the appearance of rosacea, skin blotchiness, and skinblemishes. In certain embodiments, compositions of the present inventionare applied to hair, scalp or both to improve hair health, quality andstrength, to promote hair growth or retard hair loss, to prevent ortreat dandruff, to prevent or treat seborrhea, seborrheic capitis and toimprove scalp health and moisture. In other embodiments, compositions ofthe present invention are applied to the gum, in the mouth, to preventor treat gum redness or irritation, to reduce periodontitis, to treat orprevent gingivitis, to reduce the symptoms or feeling of dry mouth. Inyet other embodiments, the compositions of the present invention areapplied to the eye to treat, prevent or reduce the appearance of red orirritated eye, to prevent or treat conjunctivitis, to improve eyemoisture, to reduce the feeling of dry eye. In other embodiments, thecompositions of the present invention are applied to the vaginal mucosato prevent or treat signs of irritation or dryness, loss of firmness.

EXAMPLES

The following test methods were used in the Examples:

Melanin Synthesis Inhibition Test

Control samples of B16(F10) murine melanoma cells were prepared andharvested as indicated below, but without addition of any test sampleand without exposure to UVB (untreated control). Other control sampleswere prepared and harvested as indicated below without addition of testsample and exposed to UVB as described below (treated control). One ormore samples of B16(F10) cells were prepared and each pre-treated with atest sample (e.g. E1) followed by UVB exposure as described below. Upontreatment, UVB stimulated melanogenesis in the cells and test compoundswere evaluated based on their ability to inhibit or slow down the rateof melanogenesis. The cells were lysed for protein measurement at 595 nmand melanin content at 470 nm. The potency of the test compounds weredetermined by comparing the % inhibition achieved by the test compoundsagainst the treated control.

Testing Procedure:

On a first day, murine melanoma B16(F10) cells were seeded in 60 mmplates with a density of ˜1 million cells per plate and incubated for 48hrs at 37° C., 5% CO₂. On day 2, the cells with a confluency rate of90-100% were treated with test compound at a predetermined concentration(e.g. 25 μg/mL) for two hours (for test compound samples only) followedby exposure to UVB 200 mJ/cm² (for test samples and treated control).The cells were harvested on day 3 (24 h post UVB irradiation for testsamples and treated control) and lysed in protein lysis buffer (50 mMTris, pH 8, 2 mM EDTA, 150 mM NaCl, and 1% Triton X 100-a nonionicsurfactant purchased from BioRad Cat.#: 161-0407), and centrifuged. Theresulting supernatant was mixed well with a protein dye assay (Bio-radprotein assay reagent) and a spectrophotometer (Molecular DevicesVERSAmax) was used to determine the optical density (protein assay OD)of the sample at 595 nm. The cell pellet remaining after removal of thesupernatant was dissolved in alkaline DMSO buffer, and the resultingsolution used for melanin absorbance assay at 470 nm to determinemelanin assay OD.

Three samples each of the untreated control, treated control, and eachtest sample were made and the Melanin and Protein OD measured for each.The normalized melanin for each untreated control (3 samples), treatedcontrol (3 samples) and test sample (3 samples for each test compound)was calculated via the following equation:Normalized Melanin=melanin assay OD/protein assay OD.

The average normalized Melanin of the untreated controls was calculated(sum of the three calculated values/3), and the average normalizedMelanin of the treated controls similarly calculated.

The Induction value of the Control was calculated via the equation:Induction value of Control=average normalized Melanin of treatedcontrol−average normalized Melanin of untreated control.

The Induction value with each test sample is then calculated via theequation:Induction value with Test Sample=normalized Melanin of the testsample−average normalized Melanin of untreated control.

The Inhibition % for each test sample is then calculated via theequation:100×[(Induction value of Control-Induction value with TestSample)/Induction value of Control].The average Inhibition % is calculated as the sum of the three resultingInhibition % values for each test sample divided by three.

The calculation sequence for % inhibition are explained by a theoreticalexample, see the following table.

Average normalized melanin 0.98 Untreated control Average normalizedmelanin 2.56 UVB treated control Induction value of control 2.56 − 0.98= 1.58 Average normalized melanin 1.04 Test sample Induction value withTest 1.04 − 0.98 = 0.06 sample Inhibition % for Test sample [(1.58 −0.06)/1.58] × 100 = 96.20%Skin Epidermal Equivalents Model as a skin Lightening Test (ΔL)

Skin epidermal equivalent tissues are available commercially fromMatTek's MelanoDerm™ System and were used for the following tests.MatTek's MelanoDerm™ System consists of normal, human-derived epidermalkeratinocytes (NHEK) and melanocytes (NHM) which have been cultured toform a multilayered, highly differentiated model of the human epidermis.Specifically, MEL-300-B tissues, each 9 mm in diameter were used in thefollowing tests.

The test materials prepared in an appropriate vehicle and testedconcentrations were applied topically to the skin model daily and theexperiment lasted for 8 days. Measurement was taken on day 9.

The macroscopic and microscopic visual tissue darkening end points weremeasured by taking pictures with a digital camera. The Degree ofLightness for each tissue (L-Value) was measured using aspectrophotometer (Konica Minolta CM-2600d). The ΔL (degree of lightnessas compared to control) for each test sample is calculated as perfollowing formula:ΔL=L-value of treated sample−L-value of control sample.Cell Viability Test

Cell Viability of the tissue during experiment was evaluated using theMTT assay described as follows. The MTT Tissue Viability Assay is acolorimetric assay system that measures the reduction of a yellowMethylthiazolyldiphenyl-tetrazolium bromide (MTT) into an insolublepurple product by the mitochondria of viable cells.

The skin epidermal tissues used previously to determine degree oflightness for each test material and of untreated tissues were used todetermine percent viable cells remaining at the end of the experiment.The skin epidermal tissues after degree of lightnes test were incubatedwith MTT reagent for 3 h. After incubation extraction buffer is added tolyse the cells and allowed to continue overnight. Samples are read usinga plate reader at a wavelength of 570 nm and compared against untreatedcontrol and expressed in % Cell Viability as of control. A reduction of≧30% cell viability as of control consider as a significant indicationof cell cytotoxicity caused by the test materials. The amount of purplecolor produced is directly proportional to the number of viable cells.

The following examples illustrates the preparation and efficacy ofPaulownia tomentosa wood extracts.

Example 1

Four extracts, at least 20 mg each, of Paulownia tomentosa were obtainedfrom Plant Extract Bank in Korea Research Institute of Biosciences &Biotechnologies representing combination of parts of the plant orindividual parts of the plant. They were derived from:stem/bark/branches combination (E1), leaves (C1), bark (C2) and stem(E2). All of the extracts were prepared using Methanol under sonicationat 50 deg C.

Example 2

The following example illustrates the preparation of Paulownia tomentosawood extract (E3) in accord with certain embodiments of the presentinvention.

Paulownia tomentosa wood powder was obtained from Kurosawa Kiri WoodSupply Shop, Kitakata-city, Japan. Ten grams (10 g) of dry wood powderwas suspended in 250 mL of reagent grade ethanol and stirred at roomtemperature for 72 h. The resulting suspension was filtered and thefiltrate dried under low pressure using rotary evaporator at 30 deg C.Dry crude extract was obtained at 3.5% yield (350 mg). The crude extractwas dissolved in methanol at a concentration of 1% and was treated withactive carbon (700 mg) for 5 min at room temperature. Suspensionfiltered through 0.45 micron filter paper. The filtrate was dried to geta visibly lighter color material, E3, 210 mg (yields 60% from crudeextract).

Example 3

The following example illustrates the preparation of Paulownia tomentosawood extract (E4) that is essentially free of β-Sitosterol and Ursolicacid.

Deionized grade water (2.4 mL) was added to the Paulownia tomentosastem/bark/branch extract (E1, 24 mg) from Example 1 above and sonicatedfor 5 min at room temperature. The resulting suspension was filteredthrough 0.45μ filter cartridge and the filtrate dried by freeze dryingto obtain water soluble components (E4; 10.3 mg) at a yield of 43%. HPLCanalysis shows the composition is essentially free from β-Sitosterol andUrsolic acid. No detectable amounts of either compounds are present inE4. The limit of detection (lod value) for β-sitosterol was 9 ppm (w/v)and for Ursolic acid was 0.5 ppm (w/v).

Example 4

The following example illustrates the skin lightening properties ofPaulownia tomentosa extracts E1-E5 and comparative examples C1-C2.

All seven extracts were tested at different concentrations of up to 2%(as listed in Table 1) via the Skin Epidermal Equivalents Model as askin Lightening Test (ΔL) as described above.

Cytotoxicity potential was determined by MTT assay for all extracts andcalculated as % reduction of cell viability as compared to control,wherein >30% reduction of cell viability constitute significantcytotoxicity issues. The results are shown in Table 1 below.

A simple one-step extraction of the wood powder with just water assolvent at room temperature was also conducted (E5) and did not resultin activity in the Skin Lightening Test. It is believed a more rigorousextraction (additional heat, agitation, etc.) may yield activity.

TABLE 1 Skin Lightening of extracts from parts of Paulownia tomentosa on3D Skin Model Degree of Conc. Lightness Std. Extract Code (%) (ΔL) Dev.E1 1 2.86 0.44 2 3.48 0.19 C1 1 none N/A 2 none N/A C2 1 * N/A 2 * N/AE2 1 2.48 0.69 2 3.71 0.43 E3 0.5 1.2  0.34 1 1.91 0.32 2 4.11 0.17 E4 22.97 0.25 E5 1 none N/A 2 none N/A 5 none N/A * Represent significantcytotoxicity issues

Example 5

The following example illustrates the Melanin Synthesis Inhibitionproperties associated with Paulownia tomentosa wood extracts.

Stem/bark/branch extract (E1) and wood powder extract (E3) were testedfor Melanogenesis Inhibition in accord with the Melanin SynthesisInhibition Test described above and also for Tyrosinase inhibition. Theresulting measurements showed that the skin lightening effect of E1 & E3were associated, at least in part, with Melanogenesis inhibition and notTryosinase inhibition. The IC50 values of extracts E1 and E3 are 30 and40 μg/mL for melanogenesis inhibition with no tyrosinase enzymeinhibition from either extracts.

Example 6 The NF-κB Inhibition Assay

The NF-κB promoter assay is conducted as follows: rat cardiac myoblastsH9c2 cells were purchased from ATCC (Manassas, Va.). Cultures weremaintained in Dulbecco's modified Eagle's medium (DMEM, Invitrogen LifeTechnologies, Carlsbad, Calif.) supplemented with 10% fetal bovineserum, 100 units/ml penicillin, and 50 ug/ml streptomycin (Invitrogenlife technologies, Carlsbad, Calif.). Typically, 1×10⁴ cells grown in96-well plates were transiently transfected with 0.45 ug total DNA perwell using Lipofectamine 2000 (Invitrogen life technologies, Carlsbad,Calif.). In all transfections, a construct with the thymidine kinasepromoter and the Renilla luciferase reporter gene (pRL-TK, Promega,Madison Wis.) was included as an internal control in addition to theluciferase promoter. One day after transfection, cells were treated withthe extracts at indicated concentrations and stimulated with 100 ng/mLof Tumor Necrosis Factor-α (TNFα, Sigma-Aldrich, St Louis, Mo.) forapproximately 24 hours before they were lysed for luciferase assays,using Dual-Luciferase Reporter System from Promega (Madison, Wis.),following manufacturer's protocol. Briefly, the firefly luciferaseactivity was measured first (representing NF-κB promoter activity),followed by the renilla luciferase (internal control), using luminometerLMAX, from Molecular Devices (Sunnyvale, Calif.). The ratio of these twoluciferase activities (RLU) was used to evaluate the activity of eachpromoter.NF-κB Inhibition=[1−(RLU _(sample) /RLU _(control))]*100Where RLU_(sample) and RLU_(control) are the normalized luciferaseactivity ratios of the sample and control, respectively.

The NF-κB inhibition assay, described above, was performed for extractsE3 and E5 in various amounts. In Tables 2a and 2b, the normalized NF-κBgene reporter activity and percent NF-κB inhibition are reported.

TABLE 2a Normalized NF-κB Percent NF-κB Gene Reporter inhibitionTreatment (Dose, as % w/v) Activity (Mean RLU) over vehicle Untreated40.5 — TNFα (100 ng/ml) 281.4 — (Stimulated) TNFα + Vehicle 244.3   0%(0.1% DMSO) TNFα + Paulownia 174.5 45.8% tomentosa (0.003%) TNFα +Paulownia 125.9 54.2% tomentosa (0.006%) TNFα + Paulownia 84.9 65.3%tomentosa (0.01%)

TABLE 2b Normalized NF-κB Percent NF-κB Gene Reporter inhibitionTreatment (Dose, as % w/v) Activity (Mean RLU) over vehicle Untreated46.9 — TNFα (100 ng/ml) 202.4 — (Stimulated) TNFα + Vehicle (0.004%161.8     0% DMSO) TNFα + Paulownia 197.7 −15.9% tomentosa water extract(0.002%) TNFα + Paulownia 201.3 −24.3% tomentosa water extract (0.004%)

Example 7 Anti-Inflammatory Effects on Release of UV-InducedPro-Inflammatory Mediators on Reconstituted Epidermis

The effect of Paulownia tomentosa extract (E3) was evaluated for topicalanti-inflammatory activity on human epidermal equivalents. Epidermalequivalents (EPI 200 HCF), multilayer and differentiated epidermisconsisting of normal human epidermal keratinocytes, were purchased fromMatTek (Ashland, Mass.). Upon receipt, epidermal equivalents wereincubated for 24 hours at 37° C. in maintenance medium withouthydrocortisone. Equivalents were topically treated (2 mg/cm²) withPaulownia tomentosa extracts in 70% ethanol/30% propylene glycol vehicle2 hours before exposure to solar ultraviolet light (1000W-Oriel solarsimulator equipped with a 1-mm Schott WG 320 filter; UV dose applied: 70kJ/m² as measured at 360 nm). Equivalents were incubated for 24 hours at37° C. with maintenance medium then supernatants were analyzed for IL-8and IL-1α cytokine release using commercially available kits (MilliporeCorp., Billerica, Mass.).

TABLE 3 Percent Inhibition Mean of IL-8 of Skin Inflamma- Treatment(Dose, as % w/v) Release (pg/mL) tion (over vehicle) Untreated, No UV170.79 — UV (60 KJ) 262.9 — UV (60 KJ) + Vehicle (70:30 253.8   0%Ethanol:Propylene Glycol) UV (60 KJ) + Paulownia 135.1 46.8% tomentosaextract 0.1% UV (60 KJ) + Paulownia 125.9 50.3% tomentosa 5%

TABLE 4 Percent Inhibition Mean of IL-1α of Skin Inflamma- Treatment(Dose, as % w/v) Release (pg/mL) tion (over vehicle) Untreated, No UV91.4 — UV (60 KJ) 188.7 — UV (60 KJ) + Vehicle (70:30 320.1   0%Ethanol:Propylene Glycol) UV (60 KJ) + Paulownia 161.2 55.2% tomentosaextract 0.1% UV(60 KJ) + Paulownia 183.3 42.7% tomentosa extract 5%

Based on the example above, topical application of the Paulowniatomentosa extract was able to significantly reduce the UV-stimulatedrelease of inflammatory mediators. Therefore, Paulownia tomentosaextracts would be expected to provide an effective anti-inflammatorybenefit when applied to skin.

Example 8 Inhibition of Reactive Oxygen Species Formation inReconstituted Epidermis

UV-induced hydrogen peroxide formation was determined using amodification of the method of Martin et al, Arch Dermatol Res. (2008)300:69-80, in reconstituted epidermis and the human epithelial cellline, KB. Epidermal equivalents (EPI 200 HCF), multilayer anddifferentiated epidermis consisting of normal human epidermalkeratinocytes, were purchased from MatTek (Ashland, Mass.). Uponreceipt, epidermal equivalents were incubated for 24 hours at 37° C. inmaintenance medium without hydrocortisone. After 24 hours, the tissueswere incubated for 30 minutes with 5 μM of the hydrogenperoxide-sensitive fluorescent probe 5-(and-6)-chloromethyl-2′,7′-dichlorodihydro-fluorescein diacetate, acetylester (CM-H2DCFDA) (Invitrogen Corp., Carlsbad, Calif.). Afterincubation, the plate was rinsed to remove excess probe and equivalentswere topically treated (2 mg/cm²) with Paulownia tomentosa extract (E3)in 70% ethanol/30% propylene glycol vehicle. The plate was immediatelyread on a fluorescent plate reader set at wavelengths 485 nmexcitation/530 nm emission to detect basal peroxide formation. The platewas then exposed to UV (1000W-Oriel solar simulator equipped with a 1 mmSchott WG 320 filter; UV dose applied 4.2 kJ/m² as measured at 360 nm).The plate was read 60 minutes post UV exposure.

TABLE 5 Mean Fluo- Percent Inhibition Treatment (Dose, as % w/v) rescentUnits of ROS Production UV + Vehicle (70:30 Ethanol: 761.5   0%Propylene Glycol) UV + Paulownia 361.4 52.5% tomentosa 0.1% UV +Paulownia 243.4 68.0 tomentosa 1.0% UV + Paulownia 261.9 65.6 tomentosa5.0%

Based on the example, topical application of Paulownia tomentosaextracts was able to significantly reduce the UV-stimulated productionof ROS in reconstituted epidermis. Therefore when applied to skin,Paulownia tomentosa extracts would be expected to provide protectionagainst induction of ROS from solar irradiation.

Example 9 Inhibition of Reactive Oxygen Species Formation in HumanEpithelial Cells

KB cells obtained from ATCC (ATCC#CCL-17, Manassas, Va.) were plated in96-well tissue culture treated plates at a density of 5000 cells/well inDulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetalbovine serum (Invitrogen Corp., San Diego, Calif.). After 48 hours,cells were incubated for 30 minutes with 5 μM of the hydrogenperoxide-sensitive fluorescent probe 5-(and-6)-chloromethyl-2′,7′-dichlorodihydro-fluorescein diacetate, acetylester (CM-H2DCFDA) (Invitrogen Corp., Carlsbad, Calif.). Afterincubation, the plate was rinsed to remove excess probe and Paulowniatomentosa extract (E3) was added at the indicated concentrations. Theplate was immediately read on a fluorescent plate reader set atwavelengths 485 nm excitation/530 nm emission to detect basal peroxideformation. The plate was then exposed to UV (1000W-Oriel solar simulatorequipped with a 1 mm Schott WG 320 filter; UV dose applied 4.2 kJ/m² asmeasured at 360 nm). The plate was read 60 minutes post UV exposure.

TABLE 6 Mean Fluo- Percent Inhibition of ROS Treatment (Dose, as % w/v)rescent Units Production (over vehicle) untreated 79.7 — UV treated220.5 — UV + Vehicle (DMSO) 219.3   0% UV + Paulownia tomentosa 160.926.6% (0.005%) UV + Paulownia tomentosa 146.2 33.3% (0.01%) UV +Paulownia tomentosa 140.2 36.1% (0.02%)

Based on the examples, treatment with Paulownia tomentosa extract wasable to significantly reduce the UV-stimulated production of ROS inhuman epithelial cells. Therefore when applied to skin, Paulowniatomentosa extracts would be expected to provide protection againstinduction of ROS from solar irradiation.

Example 10 Protection from Elastase Degradation

Human leukocyte elastase (HLE) was purchased from Sigma (St. Louis,Mo.), and reconstituted at 1 unit/ml in phosphate buffered saline (PBS,Invitrogen life Technologies, Carlsbad, Calif.). Soluble bovine neckligament elastin labeled with BODIPY FL dye was purchased from MolecularProbes, Inc. (Eugene, Or.), such that the fluorescence was quenched inthe conjugate, and could be activated upon elastase digestion. Humanleukocyte elastase (0.0625 U/ml), elastin substrate (25 μg/ml), andincreasing concentrations of test material were incubated for two hoursat 37 C. Fluorescence was measured at excitation at 490 nm and emissionat 520 nm using a fluorescent plate reader Gemini from Molecular Devices(Sunnyvale, Calif.). Background fluorescence of substrate alone had beensubtracted from each measurement. Paulownia tomentosa extract (E3) wasprepared in DMSO at a stock concentration of 10 mg/ml and seriallydiluted. Paulownia tomentosa extract inhibited HLE activity in a dosedependent manner as shown in Table 7.

TABLE 7 Paulownia tomentosa extract (Dose, as % w/v) Elastase Inhibition(%) 0 0.0 0.00001% 23.1% 0.001% 30.2% 0.005% 66.6% 0.01% 75.5% 0.02%99.4%

This example demonstrates that Paulownia tomentosa extracts can protectelastin fibers from damage and degradation.

Example 11 Inhibition of UV-Induced MMP Induction

The ability of Paulownia tomentosa extract (E3) to inhibit UV inducedmatrix metalloproteinases-1 and -9 (MMP-1 and -9) was evaluated inepidermal equivalents derived from normal human epidermal keratinocytes.MMPs are a family of enzymes that play a major role in physiologicalremodeling and pathological destruction of extracellular matrix. It iswell established that suberythemal doses of UV light induce MMPsecretion in human skin, which in turn degrades the extracellular matrixand plays a significant role in photoaging wrinkle formation and loss offirmness and elasticity. See G. J. Fisher, et al., J Investig Dermatol.Symposium Proceedings. 14(1): 20-24 (2009).

In order to evaluate the ability of Paulownia tomentosa extracts toinhibit UV induced MMPs, epidermal equivalents (EPI 200 HCF), multilayerand differentiated epidermis consisting of normal human epidermalkeratinocytes, were purchased from MatTek (Ashland, Mass.). Uponreceipt, epidermal equivalents were incubated for 24 hours at 37° C. inmaintenance medium without hydrocortisone. Equivalents were topicallytreated (2 mg/cm2) with Paulownia tomentosa extract (E3) in 70%ethanol/30% propylene glycol vehicle 2 hours before exposure to solarultraviolet light (1000W-Oriel solar simulator equipped with a 1-mmSchott WG 320 filter; UV dose applied: 70 kJ/m2 as measured at 360 nm).Equivalents were incubated for 48 hours at 37° C. with maintenancemedium then supernatants were analyzed for MMP-1 and -9 usingcommercially available kits (R&D Systems, Minneapolis, Minn.). Data inTable 8 represents the mean of 2 independent experiments, eachexperimental condition is conducted using duplicate tissues.

TABLE 8 Mean of MMP-1 Percent Inhibition Treatment (Dose, as % w/v)Release (ng/ml) of MMP-1 Production UV + Vehicle (70:30 Ethanol: 12046.2  0% Propylene Glycol) UV + Paulownia 5555.9 53.9% tomentosa 0.1% UV +Paulownia 4851.4 59.7% tomentosa 1% UV + Paulownia 4186.4 65.2%tomentosa 5%

TABLE 9 Mean of MMP-9 Percent Inhibition Treatment (Dose, as % w/v)Release (ng/ml) of MMP-9 Production UV + Vehicle (70:30 Ethanol: 20795.5  0% Propylene Glycol) UV + Paulownia 4585.9 77.9% tomentosa 0.1% UV +Paulownia 7077.2 65.9% tomentosa 5%

Based on the example topical application of Paulownia tomentosa extractwas able to significantly reduce the UV-stimulated release of MMP-1 and-9. Therefore when applied to skin, Paulownia tomentosa extracts wouldbe expected to provide protection against induction of MMP-1 and -9following solar irradiation.

Example 12 Inhibition of TNF-α-Induced MMP Induction

In order to evaluate the ability of Paulownia tomentosa extract (E3) toinhibit TNF-α induced MMPs, epidermal equivalents (EPI 200 HCF),multilayer and differentiated epidermis consisting of normal humanepidermal keratinocytes, were purchased from MatTek (Ashland, Mass.).Upon receipt, epidermal equivalents were incubated for 24 hours at 37°C. in maintenance medium without hydrocortisone. Equivalents weretopically treated (2 mg/cm2) with Paulownia tomentosa extract (E3) in70% ethanol/30% propylene glycol vehicle 2 hours before treatment withTNF-α (100 ng/mL). Equivalents were incubated for 48 hours at 37° C.with maintenance medium then supernatants were analyzed for MMP-1 and -9using commercially available kits (R&D Systems, Minneapolis, Minn.).

TABLE 10 Mean of MMP-1 Percent Inhibition Treatment (Dose, as % w/v)Release (ng/ml) of MMP-1 Production Untreated 4848.4 — TNF-α induced7867.2   0% TNF-α + Paulownia 7225.2  0.8% tomentosa 1% TNF-α +Paulownia 5370.6 31.7% tomentosa 5%

TABLE 11 Mean of MMP-9 Percent Inhibition Treatment (Dose, as % w/v)Release (ng/ml) of MMP-9 Production Untreated 13217.6 — TNF-α induced42958.6   0% TNF-α + Paulownia 35145.3 18.2% tomentosa 1% TNF-α +Paulownia 16101.1 62.5% tomentosa 5%

Based on the example topical application of Paulownia tomentosa extractwas able to significantly reduce the TNF-α stimulated release of MMP-1and -9. Therefore when applied to skin, Paulownia tomentosa extractswould be expected to provide protection against induction of MMP-1 and-9.

Example 13

The TROPOELASTIN PROMOTER ASSAY was performed using Tanacetum parthenium(parthenolide-free feverfew extract from Integrated BotanicalTechnologies of Ossining, N.Y.).

Tanacetum parthenium was diluted in cell culture media (DMEM Media ofInvitrogen, San Diego Calif.) and Paulownia tomentosa was diluted inDMSO to the concentration of “active” indicated in Table 12 below. Thecompounds were added to the transfected H9c2 cells and were incubatedfor 24 hours. Test samples were compared to respective controls. Theresults are shown in Table 12 below.

TABLE 12 Ratio of Respective Normalized Percent NFκB- Concentrations ofTropoelastin change over Inhibitor: Actives Promoter respectiveTropoelastin Compound/Extract (on active basis) Activity (RLU) controlsPromoter Untreated control — 2.69 — Tanacetum parthenium 0.002% 2.74  2%Tanacetum parthenium 0.005% 2.73  2% Vehicle control (DMSO) 0.005% 2.25— Paulownia tomentosa 0.005% 2.97 32% Paulownia tomentosa + 0.005% +0.002% 3.48 55% 2.5:1 Tanacetum parthenium Paulownia tomentosa +0.005% + 0.005% 3.64 62%   1:1 Tanacetum parthenium

As can be seen from the results shown in Table 12, Paulownia tomentosaand Tanacetum parthenium demonstrated percent changes in tropoelastinpromotion over the respective controls of 32% and 2%, respectively. Incontrast, the combination of both Paulownia tomentosa and Tanacetumparthenium demonstrated a 55% improvement in tropoelastin promotion overthe vehicle control. This was much greater than a mere additive effectin performance.

A similar synergistic effect was observed when the concentration ofTanacetum parthenium was raised from 0.002% to 0.005%. Tanacetumparthenium at the higher concentration also showed a percent change intropoelastin promotion over the control of 2%, whereas the combinationof Paulownia tomentosa and Tanacetum parthenium achieved a percentchange in tropoelastin promotion over the vehicle control of 62%.

The data demonstrates that the combination of Paulownia tomentosa and atropoelastin promoter (Tanacetum parthenium) produces a surprising andsynergistic increase in tropoelastin promotion activity.

Example 14

The following skin care composition was prepared using the ingredientsshown in Table 13 in accord with the present invention.

TABLE 13 Percentage Serial Ingredient in Formula- Number Trade NameCTFA/INCI Name tion (w/w) 1 PURIFIED WATER WATER 59.69 2 Ultrez 10Carbomer 0.60 3 VERSENE NA Disodium EDTA 0.20 4 Brij 72 Steareth-2 0.505 Brij 721 Steareth-21 1.00 6 Finsolv TN C12-15 Alkyl Benzoate 2.00 7Miglyol 812 Caprylic/Capric 2.50 Neutral Oil triglyceride 8 Emery 917Glycerin 3.00 9 PENRECO Petrolatum 0.50 SNOW WHITE 10 Dimethicone DowCorning Q7-9120 2.00 Silicone Fluide (20 cst) 11 Phenonip XBMethylparaben, ethypara- 1.00 ben, propylparaben, phenoxyethanol 12Transcutol CG EthoxyDiclycol 5.00 13 1.0% Citric Acid Citric acid 0.0114 Princess Paulownia imperialis 2.00 Tree Extract Extract 15 ButyleneGlycol Butylene Glycol 20.00 16 SODIUM HYDROX- Sodium Hydroxide Asneeded IDE PELLETS (7680-88) NF FCC Pellets Total 100.00

The above composition was prepared as follows: The purified water wasadded to a main tank at a temperature of 20-40° C. with smoothagitation. The Versene NA (disodium EDTA) was then added to the maintank. Agitation on the tank was stopped and Ultrez 10 (Carbomer) wasadded by evenly coating the top of the water mixture. The mixture wasallowed to soak and agitation and heating was started. The mixture washeated and maintained at 55-60° C., and further mixed for 15 minutes oruntil homogeneous.

An oil phase was prepared by adding Finsolv TN(C12-15 alkyl benzoate) toa clean, suitable phase container with agitation and heating to achieve55-60° C. After such temperature was achieved Brij 72 & 721 (Steareth-2,-21 resp.), Miglyol (Caprylic/Capric triglyceride), Emery 917(glycerin), and Penreco snow white (Petrolatum) were added and mixed at55-60° C. until addition to main tank.

The oil phase was added to the main tank with increased agitation andheating was stopped. The resulting mixture was mixed at high speed for10-20 minutes. At 50° C. or lower, the Dimethicone (Dow Corning SiliconeFluid) was added. The batch mixture was then cooled to 40° C. andPhenonip XB (preservative mix) was added. The mixture was further mixedfor 10 min or until uniform. Sodium hydroxide was added quickly (targetpH=5.4) with further mixing for 10 minutes or until uniform pH isachieved.

An actives premix was made by adding Transcutol CG, Butylene glycol,Citric acid, and Princess Tree extract to a separate beaker and mixingwell until uniform.

The final formulation was made by adding the actives premix to the mainphase of the main tank, and mixing for an additional 10-20 minutes todissolve completely or until uniform. The final volumes were made upwith water, the formulation mixed for 10 minutes, and pH recorded.

The samples of composition were placed in 50° C. oven for 2 week andshowed primary good stability.

Example 15

The following skin care composition was prepared using the ingredientsshown in Table 14 in accord with the present invention.

TABLE 14 Serial Percentage in Number CTFA/INCI Name Formulation (w/w) 1Purified Water 75.55 2 Disodium EDTA 0.15 3 AmmoniumAcryloyldimethyltaurate/VP 0.30 Copolymer 4 Chlorphenesin C 0.20 5Butylene glycol 6.00 6 Cetearyl Olivate/Sorbitan Olivate 0.50 7 StearicAcid 0.50 8 Ethylhexylglycerin 1.00 9 Cyclopentasiloxne &Cyclohexasiloxane 5.00 10 Cyclopentasiloxane & Dimethicone 3.00Crosspolymer 11 Dimethiconol & Dimethicone 2.00 12 Sodium hydroxide 2.4013 Polyacrylate 13 & Polyisobutene & 1.00 Polysorbate 20 14Methylisothiazolinone 0.15 15 Fragrance 0.01 16 FD&C Red 0.12 17 D&CYellow 0.12 18 Paulownia imperialis 2.00 (Princess Tree) Extract Total100.00

The above composition was prepared as follows: The purified water wasadded to a main tank followed by addition of disodium EDTA and mixeduntil the EDTA dissolved. Ammonium acryloyldimethyltaurate/VP Copolymerwas sprinkled in and the resulting mixture heated to 70-75° C. After settemperature is achieved, Cetearyl Olivate/Sorbitan Olivate and Stearicacid were added while agitating the mixture for 5 minutes at settemperature.

An actives premix was prepared by dissolving Princess tree extract intobutylene glycol at 40-50° C. in a separate container. Polyacrylate 13and polyisobutene and polysorbate 20 were added to the Princess tree mixand then mixed until uniform. The temperature was lowered to 35-40° C.and set aside until ready to add to the main tank.

A Chlorophenesin premix was prepared by adding Chlorophehesin tobutylenes glycol in a separate container and heating to 35° C., followedby addition of Methylisothiazolinone. The resulting mixture was heatedto 50-55° C. and the temperature maintained until ready to mix into maintank.

An oil phase was prepared in a separate container by addingCyclopentasiloxane and Dimethicone Crosspolymer to Cyclopentasiloxaneand Cyclohexasiloxane while mixing and heating to 55-60° C. untiluniform. Then Ethylhexylglycerin was added and mixed until uniform,followed by addition of Stearic acid with the temperature maintainedbetween 55-60° C.

The oil phase was then added to the main tank slowly with vigorousagitation at a temperature of 70-75° C. Dimethiconol and dimethiconewere added to the main batch and the resulting mixture stirred for 20minutes or until uniform, after which the heating was stopped. The mainpH was adjusted to between 5.0-5.5 with sodium hydroxide. TheChlorophenesin phase was added slowly at 50-55° C. while maintainingagitation. The mixture was cooled to 35-40° C. and the fragrance, FD&Cred and D&C Yellow added and mixed well while maintaining thetemperature.

The final formulation was achieved by adding the active premix to themain tank slowly with gentle mixing. The mixture was mixed for anadditional 10-20 until uniform. The final volumes were made up withwater, the formulation mixed for 10 minutes, and pH recorded.

The samples of composition were placed in 50° C. oven for 2 week andshowed primary good stability.

What is claimed is:
 1. A method of improving a sign of aging in skinselected from the group consisting of improving skin firmness, improvingskin texture, improving the appearance of wrinkles, treating externalaggression in skin, or combinations thereof, said method comprisingtopically applying to human skin in need of improving the sign of agingin skin a therapeutically effective amount of an extract of Paulowniatomentosa wood containing an effective amount of paulownin to said humanskin.
 2. The method of claim 1, wherein said extract is obtained withone or more solvents, wherein one or more solvents are selected fromC₁-C₈ alcohols, C1-C8 glycols.
 3. The method of claim 1, wherein saidextract is obtained with one or more solvents selected from ethanol,methanol, or a combinations thereof.
 4. The method of claim 1, whereinsaid extract is obtained with a solvent having a dielectric constant offrom about 4 to about 60 at 20° C.
 5. The method of claim 1, whereinsaid extract of Paulownia tomentosa wood containing paulownin is appliedin an amount of greater than zero to about 20% to skin in need ofimproving a sign of aging.
 6. The method of claim 1, wherein saidextract of Paulownia tomentosa wood containing paulownin is applied inan amount of from about 0.01 to about 5% to skin in need of improving asign of aging.
 7. The method of claim 1, wherein said said extract ofPaulownia tomentosa wood containing paulownin is administered in acomposition, wherein said composition comprises the effective amount ofthe extract of Paulownia tomentosa wood containing paulownin and acarrier, wherein said composition is administered to skin in need ofimproving a sign of aging, wherein said composition is the form of asolution, suspension, lotion, cream, serum, gel, stick, spray, ointment,liquid wash, soap bar, shampoo, hair conditioner, paste, foam, powder,mousse, shaving cream, hydrogel, or film-forming product.
 8. The methodof claim 7, wherein said composition further comprises a tropoelastinpromoter.
 9. The method of claim 8, wherein said tropoelastin promoteris selected from the group consisting of blackberry extracts, cotinusextracts, feverfew extracts, extracts of Phyllanthus niruri andcombinations of two or more thereof.
 10. The method of claim 7, whereinsaid composition further comprises a collagen promoter.
 11. The methodof claim 10, wherein the collagen promoter is a non-retinoid collagenpromoter.
 12. The method of claim 10, wherein the non-retinoid collagenpromoter is selected from the group consisting of extracts of Tanacetumparthenium, extracts of Centella asiatica, extracts of Siegesbeckiaorientalis, extracts of soy, and combinations of two or more thereof.13. The method of claim 10, wherein the collagen promoter is a retinoid.14. The method of claim 13, wherein the retinoid is selected from thegroup consisting retinol, retinal, retinyl acetate, retinyl propionate,retinyl linoleate, retinyl palmitate, and combinations of two or morethereof.
 15. The method of claim 1, wherein improving a sign of agingcomprises improving improving skin firmness comprising topicallyapplying to human skin in need of improving skin firmness an effectiveamount of an extract of Paulownia tomentosa wood containing paulownin.16. The method of claim 15, wherein said human skin in need of improvingskin firmness comprises skin that is sagging, loose, lax, rough,wrinkly, thinned, uneven, or a combination of two or more thereof. 17.The method of claim 1, wherein improving a sign of aging comprisesimproving the appearance of wrinkles in the skin comprising applying tohuman skin in need of improving the appearance of wrinkles in the skinan effective amount of the extract of Paulownia tomentosa woodcontaining paulownin.
 18. The method of claim 15, wherein said humanskin in need of improving the appearance of wrinkles in the skincomprises skin having wrinkles, fine lines, or a combination thereof.19. The method of claim 7, wherein said composition further comprisesbutylene glycol.